EA044940B1 - DERIVATIVES OF CONDENSED TRICYCLIC RING AS PHOSPHATASE INHIBITORS WITH SRC2 HOMOLOGY DOMAIN - Google Patents

Description

Disclosed herein are certain fused tricyclic ring derivatives that are Src2 homology domain phosphatase (SHP2) inhibitors and therefore can be used to treat diseases that can be treated by inhibition of SHP2. Also presented are pharmaceutical compositions containing such compounds and methods for preparing such compounds.

State of the art

SHP2 is a non-receptor protein phosphatase ubiquitously expressed in a variety of tissues and cell types (for reviews, see: Tajan Met al., Eur J Med Genet 2016 58(10):509-25; Grossmann KS et al., Adv Cancer Res 2010 106: 53-89). SHP2 consists of two Src homology 2 domains (N-SH2 and C-SH2) at its NH2 terminus, a catalytic PTP (protein tyrosine phosphatase) domain, and a C-terminal tail with regulatory properties. In the ground state, intermolecular interactions between the SH2 domains and the PTP domain prevent substrates from entering the catalytic pocket, maintaining SHP2 in a closed, self-inhibited conformation. In response to stimulation, SHP2 activating proteins bearing phosphotyrosine motifs bind to SH2 domains, resulting in active site targeting and enzymatic activation of SHP2.

SHP2 plays important roles in fundamental cellular functions, including proliferation, differentiation, cell cycle maintenance, and motility. Through dephosphorylation of its associated signaling molecules, SHP2 regulates multiple intracellular signaling pathways in response to a wide range of growth factors, cytokines and hormones. Cell signaling processes in which SHP2 is involved include the RAS-MAPK (mitogen-activated protein kinase), PI3K (phosphoinositol 3-kinase)-AKT and AK-STAT pathways.

The RAS-MAPK signaling pathway is critical for tumor formation and maintenance. Genes encoding various components of this pathway, including RTK (receptor tyrosine kinases), SHP2, NF1, RAS or RAF are mutated in cancer, leading to upregulation of MAPK signaling. SHP2 also plays a signal-enhancing role in this pathway, acting downstream of RTK and upstream of RAS. It has been demonstrated that the survival of RTK-driven cancer cells is dependent on SHP2. Thus, SHP2 inhibitor has been proposed as a valuable treatment for RTK-driven cancers (see Prahallad, A. et al. Cell Reports 12, 1978-1985 (2015); Chen YN, Nature 535, 148-152 (2016)).

Much effort has been devoted to developing pharmacological agents targeting various nodes in the RAS-MAPK pathway, such as RTK inhibitors, BRAF inhibitors, and MEK inhibitors for the treatment of cancer. Although these agents show good initial efficacy, resistance to them often occurs. One common mechanism of resistance involves the activation of RTKs, which fuel the reactivation of MAPK signaling. Because SHP2 is required downstream of multiple RTKs for signal transduction, inhibition of SHP2 may provide a general strategy for preventing resistance to anticancer drugs targeting the MAPK pathway. Recent studies in preclinical models have shown that SHP2 inhibition overcomes resistance and offers synergistic therapeutic effects when combined with an ALK inhibitor (see Dardaei L et al. Nat Med. 24, 512-17 (2018)), a MEK inhibitor (see Mainardi, S. et al. Nat. Med. https://doi.org/10.1038/s41591-018-0023-9 (2018); Ruess, D. A. et al. Nat. Med. https://doi.org/10.1038/ s41591018-0024-8 (2018); Wong, G. S. et al. Nat. Med. https://doi.org/10.1038/s41591-018-0022-x (2018); Fedele C et al. Cancer Discov pii: CD -18-0444.doi: 10.1158/2159-8290.CD-18-0444 (2018)) or a BRAF inhibitor (see Prahallad, A. et al. Cell Reports 12, 1978-1985 (2015)). Notably, combined MEK/SHP2 inhibition has been identified as having potential for the treatment of KRAS-driven cancers with most commonly mutated oncogenes. Despite years of effort, inhibitors directly targeting KRAS have not yet been successfully developed for clinical use. Inhibition of MEK, a downstream effector of KRAS, only transiently suppressed MAPK signaling. The discovery of dual MEK/SHP2 inhibition takes important steps in long-term efforts to better understand and therapeutically target KRAS-driven cancers.

Given the vital physiological functions that SHP2 performs, targeted deregulation of SHP2 is expected to have broad therapeutic applications. Gain of functional mutations in PTPN11, the gene encoding SHP2, have been causally associated with several human diseases, including Noonan syndrome, juvenile myelomonocytic leukemia, acute myeloid leukemia, myelodysplastic syndrome, and B-lymphoblastic acute lymphoblastic leukemia. SHP2 functions as an oncogene, and its overexpression and/or activating mutations have been described in various solid tumors.

- 1,044,940 tumors such as neuroblastoma, breast cancer, colon cancer, lung cancer, melanoma and hepatocellular carcinoma.

In addition, SHP-2 is thought to mediate inhibitory signaling of multiple receptor immune checkpoints (e.g., PD-1) through dephosphorylation of CD28. To support this notion, dominant negative SHP-2 abolishes PD-1 signaling and restores cytotoxic CAR T cell function. Therefore, SHP-2 inhibitors have potential for use in combination therapy with existing targeted and immuno-oncology (IO) agents.

In addition to human tumors, increased SHP2 expression or activity has been implicated in the pathogenesis of autoimmune diseases such as systemic lupus erythematosus (Wang J et al. J Clin Invest. 2016 Jun 1; 126(6):2077-92) and rheumatoid arthritis ( see Stanford S. M et al. Arthritis Rheum. 2013 May; 65(5): 1171-80; Maeshima K et al. JCI Insight. 2016 May 19; 1(7)). Recently, SHP2 has also been characterized as a molecular checkpoint for TGFP-induced JAK2/STAT3 signaling, suggesting that inhibition of SHP2 may offer therapeutic benefit for the treatment of fibrosis (see Zehender A et al. Nat Commun. 2018 Aug 14;9( 1):3259). Therefore, SHP2 represents a very attractive target for the development of new therapies to treat various diseases.

The essence of the invention

In the first aspect, a compound of formula (IIB) is presented:

(PV) or a pharmaceutically acceptable salt thereof, where:

where Z is -Y-M;

Y is a bond;

M is alkyl substituted with -O(alkylene)OR ^a where R ^a is alkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl or heterocyclylalkyl; or

Z is methoxymethyloxy, methoxyethyloxy, methoxymethyl, cyclopropylmethyloxymethyl or oxetan-3-ylmethyloxymethyl;

or

Z is (methoxymethoxy)methoxymethyl;

R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen, alkyl or halogen;

R ⁵ and R ⁶ are independently selected from hydrogen, alkyl or halogen;

L is a bond, O, S, S(O) or S(O) ₂ ;

R ⁹ is hydrogen, alkyl, halogen or amino;

R ¹⁰ is hydrogen, alkyl, halogen, hydroxy, hydroxyalkyl or -CD2OH; And

R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

In a fifth aspect, there is provided a pharmaceutical composition comprising a compound of formula (IIB) (or any of the embodiments thereof described herein) or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable excipient.

In a sixth aspect, there is provided a method of treating a disease treatable by inhibition of SHP2 in a patient, wherein the method comprises administering to the patient, preferably a patient in need of such treatment, a therapeutically effective amount of a compound of formula (IIB) (or any of the embodiments described herein) or involves administering to a patient, preferably

- 2 044940 to a patient in need of such treatment, a pharmaceutical composition containing a compound of formula (IIB) (or any of the embodiments described herein) and a pharmaceutically acceptable excipient. In one embodiment, the disease is cancer. In another embodiment, the disease is a cancer selected from lung, stomach, liver, colon, kidney, breast, pancreas, juvenile myelomonocytic leukemia, neuroblastoma, melanoma and acute myeloid leukemia. In one embodiment, the disease is selected from Noonan syndrome and Leopard syndrome.

In a seventh aspect, there is provided a compound of formula (IIB) (or any of the embodiments described herein) or a pharmaceutically acceptable salt thereof for use as a drug.

In an eighth aspect, there is provided the use of a compound of formula (IIB) or a pharmaceutically acceptable salt thereof (or any of the embodiments described herein) in the manufacture of a medicament for the treatment of a disease in a patient in need of such treatment in which SHP2 activity contributes to pathology and/or symptoms of the disease.

In a ninth aspect, a method of inhibiting SHP2 is provided, wherein the method comprises contacting SHP2 with a compound of formula (IIB) (or any of the embodiments described herein) or a pharmaceutically acceptable salt thereof; or contacting SHP2 with a pharmaceutical composition containing a compound herein (or any of the embodiments described herein) or a pharmaceutically acceptable salt and a pharmaceutically acceptable excipient thereof.

Embodiments of the embodiment are provided herein below.

Detailed description

Some structures presented in this document are drawn with one or more floating substituents. Unless otherwise stated or otherwise clear from the context, substituent(s) may be present on any atom of the ring to which it is attached if chemically possible and valence rules permit. For example, in the structure:

R ⁹

R ¹ >=N /—< _R u ^R M°\

R3 =Л )=/ N-к '— ^{/ R N} аж ^Rl ° Jk NW R ⁶

Z r4 r5

R ³ and R ⁴ substituents can replace any hydrogen on the 5-membered ring.

Definitions.

Unless otherwise specified, the following terms used in the description and claims are defined for the purposes of this invention and have the following meaning.

Alkyl means a linear saturated monovalent hydrocarbon radical containing from one to six carbon atoms, or a branched saturated monovalent hydrocarbon radical containing from three to six carbon atoms, for example, methyl, ethyl, propyl, 2-propyl, butyl, pentyl and the like. One skilled in the art will appreciate that the term alkyl can include alkylene groups.

Alkylene means a linear saturated divalent hydrocarbon radical containing from one to six carbon atoms, or a branched saturated divalent hydrocarbon radical containing from three to six carbon atoms, unless otherwise specified, for example, methylene, ethylene, propylene, 1-methylpropylene, 2- methylpropylene, butylene, pentylene and the like.

Alkenyl means a linear monovalent hydrocarbon radical containing from two to six carbon atoms or a branched monovalent hydrocarbon radical containing from three to six carbon atoms containing a double bond, for example propenyl, butenyl and the like.

Alkyldienyl means alkenyl, as defined above, which is attached via a terminal divalent carbon.

the alkyldienyl group is enclosed in a frame, which is indicated by an arrow.

Alkylthio means the radical -SR, where R is alkyl as defined above, for example methylthio, ethylthio and the like.

Alkylsulfonyl means -SO2R radical, where R is alkyl as defined above, for example, methylsulfonyl, ethylsulfonyl and the like.

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Alkyl sulfoxide means -SOR radical, where R is alkyl as defined above, for example, methyl sulfoxide, ethyl sulfoxide and the like.

Amino means -NH2.

Alkylamino means -NHR radical, where R is alkyl as defined above, for example, methylamino, ethylamino, propylamino or 2-propylamino and the like.

Acylamino means -NHC(O)R radical, where R is alkyl as defined above, for example, acetylamino, propionylamino and the like.

Aminoalkyl means a linear monovalent hydrocarbon radical containing from one to six carbon atoms, or a branched monovalent hydrocarbon radical containing from three to six carbon atoms, substituted -NR'R, where R' and R are independently hydrogen, alkyl, haloalkyl, hydroxyalkyl or alkoxyalkyl, or R and R together with the nitrogen atom to which they are attached form an optionally substituted heterocyclyl, each of which is defined herein, for example, aminomethyl, aminoethyl, methylaminomethyl, morpholinylethyl, piperazin-1-ylethyl and the like.

Alkoxy means the radical -OR, where R is alkyl as defined above, for example methoxy, ethoxy, propoxy or 2-propoxy, n-, iso- or tert-butoxy and the like.

Alkoxyalkyl means a linear monovalent hydrocarbon radical containing from one to six carbon atoms, or a branched monovalent hydrocarbon radical containing from three to six carbon atoms, substituted by at least one alkoxy group, such as one or two alkoxy groups as defined above, for example , 2-methoxyethyl, 1-, 2- or 3-methoxypropyl, 2-ethoxyethyl and the like.

Alkoxycarbonyl means a -C(O)OR radical where R is alkyl as defined above, eg methoxycarbonyl, ethoxycarbonyl and the like.

Aryl means a monovalent monocyclic or bicyclic aromatic hydrocarbon radical containing from 6 to 10 ring atoms, for example, phenyl or naphthyl.

Aralkyl means -(alkylene)^ where R is aryl as defined above, for example benzyl or phenethyl.

Cycloalkyl means a monocyclic saturated monovalent hydrocarbon radical containing from three to ten carbon atoms, optionally substituted by one or two substituents independently selected from alkyl, halogen, alkoxy, hydroxy and cyano, unless otherwise indicated. Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and the like.

Cycloalkylalkyl means -(alkylene)-R radical, where R is cycloalkyl as defined above, for example, cyclopropylmethyl, cyclohexylmethyl and the like.

Cycloalkylene means, unless otherwise specified, a monocyclic saturated divalent hydrocarbon radical containing from three to six carbon atoms, optionally substituted by one or two substituents independently selected from alkyl, halogen, alkoxy, hydroxy and cyano, each of which is defined herein. Examples include, but are not limited to, cyclopropylene, cyclobutylene, cyclopentylene or cyclohexylene, and the like.

Carboxy means -C(O)OH.

Dialkylamino means -NRR' radical, where R and R' are alkyl as defined above, for example, dimethylamino, methylethylamino and the like.

Aminosulfonyl means a -SO2NRR' radical wherein R and R' are independently hydrogen, alkyl, haloalkyl, hydroxyalkyl or alkoxyalkyl, each as defined herein, for example, aminosulfonyl, methylaminosulfonyl, dimethylaminosulfonyl and the like.

Aminocarbonyl means a -CONRR' radical, wherein R and R' are independently hydrogen, alkyl, haloalkyl, hydroxyalkyl or alkoxyalkyl, each as defined herein, for example, aminocarbonyl, methylaminocarbonyl, dimethylaminocarbonyl, ethylmethylaminocarbonyl and the like.

Aminocarboxy means a -C(O)ONRR' radical, wherein R and R' are independently hydrogen, alkyl, haloalkyl, hydroxyalkyl or alkoxyalkyl, each of which is defined herein, for example, aminocarbonyloxy, methylaminocarbonyloxy, dimethylaminocarbonyloxy and the like.

Halogen means fluorine, chlorine, bromine or iodine, preferably fluorine or chlorine.

Haloalkyl means an alkyl radical as defined above which is substituted with one or more halogen atoms, for example one to five halogen atoms such as fluorine or chlorine, including substituted with various halogens, for example -CH ₂ Cl, -CF ₃ , -CHF2 , -CH2CF3, -CF2CF3, -CF(CH ₃ )2 and the like. If the alkyl is replaced only by fluorine, it may be called fluoroalkyl in the present invention.

Haloalkoxy means -OR radical, where R is haloalkyl as defined above, for example -OCF ₃ , -OCHF ₂ and the like. If R is haloalkyl, where the alkyl is substituted only by fluorine, it may be called fluoroalkoxy in the present invention.

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Hydroxyalkyl means a linear monovalent hydrocarbon radical containing from one to six carbon atoms, or a branched monovalent hydrocarbon radical containing from three to six carbon atoms, substituted by one or two hydroxy groups, provided that if two hydroxy groups are present, they are not on the same or the same carbon atom. Typical examples include, but are not limited to, hydroxymethyl, 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, 1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl, 3-hydroxybutyl, 4hydroxybutyl, 2,3-dihydroxypropyl, 1-( hydroxymethyl)-2-hydroxyethyl, 2,3-dihydroxybutyl, 3,4dihydroxybutyl and 2-(hydroxymethyl)-3-hydroxypropyl, preferably 2-hydroxyethyl, 2,3dihydroxypropyl and 1-(hydroxymethyl)-2-hydroxyethyl.

Heterocyclyl means a saturated or unsaturated monovalent monocyclic or bicyclic ring containing 4-10 ring atoms, in which one, two or three ring atoms are heteroatoms selected from N, O and S(O) _n , where n is an integer from 0 to 2, the remaining ring atoms are C. Additionally, one or two ring carbon atoms in the heterocyclic ring may be optionally substituted with a -CO- group. More specifically, the term heterocyclyl includes, but is not limited to, pyrrolidino, piperidino, homopiperidino, 2-oxopyrrolidinyl, 2oxopiperidinyl, morpholino, piperazino, tetrahydropyranyl, thiomorpholino, 6,7,8,9-tetrahydro-4Hpyrido[1,2-a] pyrimidin-4-one, 6,7-dihydropyrimido[2,1-c][1,4]oxazin-4(9H)-one and the like. When the heterocyclyl ring is unsaturated, it may contain one or two ring double bonds, provided that the ring is not aromatic. When a heterocyclyl group contains at least one nitrogen atom, it is also referred to herein as heterocycloamino and is a subset of the heterocyclyl group.

Heterocyclylene means, unless otherwise specified, a saturated or unsaturated divalent monocyclic or bicyclic ring containing 4-6 ring atoms, in which one, two or three ring atoms are heteroatoms selected from N, O and S(O)n, where n is an integer from 0 to 2, the remaining ring atoms being C. The heterocyclylene may be optionally substituted with one or two substituents independently selected from alkyl, halogen, haloalkyl, haloalkoxy, cyano or hydroxy, each of which is defined herein.

Heteroaryl means a monovalent monocyclic or bicyclic aromatic radical containing 5-10 ring atoms, unless otherwise specified, where one or more (in one embodiment, one, two or three) ring atoms is a heteroatom selected from N, O or S, the remaining ring atoms are carbon. Typical examples include, but are not limited to, pyrrolyl, thienyl, thiazolyl, imidazolyl, furanyl, indolyl, isoindolyl, oxazolyl, isoxazolyl, benzothiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazolyl, tetrazolyl, and the like. As defined herein, the terms heteroaryl and aryl are mutually exclusive. When a heteroaryl ring contains 5 or 6 ring atoms, it is also referred to herein as a 5- or 6-membered heteroaryl.

Heteroaralkyl means -(alkulene)-R, where R is heteroaryl as defined above, for example benzyl or phenethyl.

The term oxo as used herein, alone or in combination, refers to =(O).

If necessary, any definition herein may be used in combination with any other definition to describe a constituent structural group. By convention, the final element of any such definition is what is attached to the parent group. For example, the alkoxyalkyl moiety means that the alkoxy group is attached to the parent molecule through an alkyl group.

A pharmaceutically acceptable salt of a compound of formula (IIB) means a salt that is pharmaceutically acceptable and that has the desired pharmacological activity of the parent compound. Such salts include acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like; or formed with organic acids such as formic acid, acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2naphthalenesulfonic acid, 4 -toluenesulfonic acid, camphorsulfonic acid, glucoheptonic acid, 4,4'-methylenebis-(3-hydroxy-2-en-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid and the like; or salts formed when an acidic proton present in the parent compound is either replaced by a metal ion, for example an alkali metal ion, an alkaline earth metal ion or an aluminum ion; or coordinates with an organic base such as

- 5 044940 ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine and the like. It is understood that the pharmaceutically acceptable salts are non-toxic. Additional information regarding suitable pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, PA, 1985, which is incorporated herein by reference in its entirety.

The compounds of formula (IIB) may have asymmetric centers. Compounds of formula (IIB) containing an asymmetrically substituted atom can be isolated in optically active or racemic forms. Individual stereoisomers of compounds can be prepared synthetically from commercially available starting materials that contain chiral centers, or by preparing mixtures of enantiomeric products followed by resolution, such as conversion to a mixture of diastereomers followed by separation or recrystallization, chromatographic methods, direct separation of enantiomers on chiral chromatography columns or any other suitable method known in the art. All chiral, diastereomeric, all mixtures of chiral or diastereomeric forms and racemic forms are included within the scope of this specification unless a specific stereochemistry or isomeric form is specifically stated. One skilled in the art will also appreciate that when a compound is designated as an (R) stereoisomer, it may contain the corresponding (S) stereoisomer as an impurity and vice versa.

Some compounds of formula (IIB) may exist as tautomers and/or geometric isomers. All possible tautomers and cis- and trans-isomers, in individual forms and mixtures thereof, are included within the scope of the present description. Additionally, as used herein, the term alkyl includes all possible isomeric forms of said alkyl group, although only a few examples are given. In addition, when cyclic groups such as aryl, heteroaryl, heterocyclyl are substituted, they include all positional isomers, although only a few examples are given. In addition, all hydrates of the compound of formula (IIB) are included within the scope of the present description.

Compounds of formula (IIB) may also contain non-naturally occurring amounts of isotopes on one or more of the atoms that make up such compounds. Non-naturally occurring amounts of an isotope can be defined as those ranging from the naturally occurring amount to the amount that constitutes 100% of the atom in question, which differ only in the presence of one or more isotopically enriched atoms. Exemplary isotopes that may be included in the compounds of the present invention, such as a compound of formula (IIB) (and any embodiment thereof described herein, including specific compounds), include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur , fluorine, chlorine and iodine, such as ² H, ³ H, ⁿ C, ¹³ C, ¹⁴ C, ¹³ N, ¹⁵ N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ^{32 R, 33 R} , ³⁵ S, ¹⁸ F , ³⁶ Cl, ¹²³ I and ¹²⁵ I, respectively. Isotopically labeled compounds (eg, ³ H and ¹⁴ C labeled) can be used in compound or substrate tissue distribution assays. Tertiated (i.e. ^3H ) and carbon-14 (i.e. ^14C ) isotopes can be used due to their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium (ie, ^2H ) may provide certain therapeutic benefits derived from greater metabolic stability (eg, increased in vivo half-life or reduced dosage requirements). In some embodiments, the compounds described herein, including those listed in Table. 1 below, one or more hydrogen atoms are replaced by ² H or ³ H, or one or more carbon atoms are replaced by ¹³ C- or ¹⁴ C-rich carbon. Positron-active isotopes such as ¹⁵ O, ¹³ N, ⁿ C and ¹⁵ F are used in positron emission tomography (PET) studies to test for substrate receptor occupancy. Isotopically labeled compounds can generally be prepared by the following methods, similar to those described in the schemes or examples herein, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

Optionally substituted aryl means aryl that is optionally substituted with one, two or three substituents independently selected from alkyl, hydroxyl, cycloalkyl, carboxy, alkoxycarbonyl, hydroxy, alkoxy, alkylthio, alkylsulfonyl, amino, alkylamino, dialkylamino, halogen, haloalkyl, haloalkoxy and cyano .

Optionally substituted aralkyl means -(alkylene)-R, where R is optionally substituted aryl as defined above.

Optionally substituted heteroaryl means heteroaryl as defined above which is optionally substituted with one, two or three substituents independently selected from alkyl, alkylthio, alkylsulfonyl, hydroxyl, cycloalkyl, carboxy, alkoxycarbonyl, hydroxy, alkoxy, halogen, haloalkyl, haloalkoxy, amino, alkylamino , dialkylamino and cyano.

Optionally substituted heteroaralkyl means -(αalkylene)-R, where R is optionally substituted heteroaryl as defined above.

Optionally substituted heterocyclyl means a heterocyclyl as defined above that is optionally substituted with one, two or three substituents independently selected from alkyl, alkylthio, alkylsulfonyl, hydroxyl, cycloalkyl, carboxy, alkoxycarbonyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, aminoalkyl, halogen, haloalkyl , haloalkoxy and cyano.

Optionally substituted heterocyclylalkyl means -(alkylene)-R, where R is an optionally substituted heterocyclyl as defined above.

Pharmaceutically acceptable carrier or excipient means a carrier or excipient that is used in the preparation of a pharmaceutical composition that is generally safe, non-toxic, and not biologically or otherwise undesirable, and includes a carrier or excipient acceptable for veterinary use as, as well as pharmaceutical use in humans. The pharmaceutically acceptable carrier/excipient used in the specification and claims includes one or more such excipients.

The term approximately, as used herein, is intended to refer to the numerical values it changes, designating such value as variable within error. When specific error limits are not stated, such as standard deviation from the mean value reported on a graph or data table, the term approximately should be understood to mean a range that will cover ±10%, preferably ±5%, of the stated value and range, inclusive .

The term disease as used herein is generally synonymous with and is used interchangeably with the terms disorder, syndrome and condition (as in medical condition), since all of them reflect an abnormal condition of the human or animal body or one of its parts that interferes with normal functioning, usually manifests itself in the form of distinctive signs and symptoms and results in a reduction in the length or quality of life of a person or animal.

The term combination therapy means the administration of two or more therapeutic agents to treat a disease or disorder described herein. Such administration involves coadministration of these therapeutic agents in a substantially simultaneous manner, for example, in a single capsule with a fixed ratio of active ingredients or in multiple separate capsules for each active ingredient. In addition, such administration also includes sequential application of each type of therapeutic agent. In either case, the treatment regimen will provide the beneficial effects of the drug combination in treating the conditions or disorders described herein.

The term patient is generally synonymous with the term subject and includes all mammals, including humans. Examples of patients include humans, livestock such as cows, goats, sheep, pigs and rabbits, and companion animals such as dogs, cats, rabbits and horses. Preferably, the patient is a human.

Treatment or treatment of a disease includes:

(1) disease prevention, i.e. preventing the development of clinical symptoms of a disease in a mammal that may be susceptible or predisposed to the disease, but is not yet experiencing or exhibiting symptoms of the disease;

(2) inhibition of the disease, i.e. stopping or reducing the development of the disease or its clinical symptoms; or (3) alleviation of the disease, i.e. causing regression of the disease or its clinical symptoms. In one embodiment, treat or treat means (2) or (3) above.

A therapeutically effective amount means an amount of a compound of formula (IIB) or a pharmaceutically acceptable salt thereof which, when administered to a patient to treat a disease, is sufficient to effect such treatment of the disease. The therapeutically effective amount will vary depending on the compound, the disease and its severity and age, body weight, etc. mammal being treated.

The terms inhibition and reduction or any modification of these terms with respect to SHP2 include any measurable reduction or complete inhibition to achieve the desired result. For example, there may be a reduction of about, at most, about, or at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55% , 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or more, or any range derived herein, of a decrease in SHP2 activity compared to normal.

Typical compounds of formula (IIB) are described in table. 1 below.

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Table 1

Connection No. Structure Name 14 (-SS-L \ ΥΎ g № γ Ν > η ₂ ν^\ (3 -((3S,4 8)-4-amino-3-methyl-2oxa-8-azaspiro[4.5]decan-8yl)-6-(((6a8,88)-8(methoxymethoxy)-ba, 7,8,9tetrahydro-6H-pyrido[3,2b]pyrrolo[1,2-d][1,4]oxazin-4yl)thio)pyrazin-2-yl)methanol 15 / VnX^s^ \ ϊΎ Γ f / NX, νη ₂ meo γ ν η _? ^ί no / ' ^-ο (3 -((3 S,4 8)-4-amino-3 -methyl-2oxa-8-azaspiro[4.5]decan-8yl)-6-(((6a8,8 8)-8-(2methoxyethoxy)- 6a, 7,8,9tetrahydro-6H-pyrido[3,2b]pyrrolo[1,2-d][1,4]oxazin-4yl)thio)pyrazin-2-yl)methanol 33 Ύ* ω U k (3 -((3 S,4 8)-4-amino-3 -methyl-2oxa-8-azaspiro[4.5]decan-8yl)-6-(((6a8,88)-8- ((cyclopropylmethoxy)methyl)6a,7,8,9-tetrahydro-6Hpyrido[3,2-b]pyrrolo[1,2d][l,4]oKca3HH-4- yl)thio)pyrazin-2-yl)methanol 34 0 'ί uh co With Ho (3 -((3 S,4 8)-4-amino-3 -methyl-2oxa-8-azaspiro[4.5]decan-8yl)-6-(((6a8,8 8)-8-((oxetan- 3 ylmethoxy)methyl)-6a, 7,8,9tetrahydro-6H-pyrido[3,2-

- 8 044940

b]pyrrolo[1,2-d][1,4]oxazin-4yl)thio)pyrazin-2-yl)methanol 38 z τ / 0’ (3 -((8)-5-amino-13-oxa-9azadispiro[3.1,56.24]tridecane9-nn)-6-(((6aS,8S)-8(methoxymethyl)-ba, 7,8,9tetrahydro- 6H-pyrido[3,2b]pyrrolo[1,2-d] [1,4]oxazin-4yl)thio)pyrazin-2-yl)methanol 39 /^Ύ^Ο \ ΊΓη / ^Ν -χ<0 NH, MeO γ N > - ² NSU X> ^-0 (3 -((8)-5-amino-13-oxa-9azadispiro[3.1,56.24]tridecan9-yl)-6-(((6а8,88)-8((methoxymethoxy)methyl)6а,7,8, 9-tetrahydro-6Hpyrido[3,2-b]pyrrolo[1,2d][l,4]oKca3HH-4yl)thio)pyrazin-2-yl)methanol 40 O.. I\*2\/w ω oJ^t °°2 s (3-((3 8.48)-4-amino-3-methyl-2oxa-8-azaspiro[4.5]decan-8yl)-6-(((6a8.88)-8- (((methoxymethoxy)methoxy) methyl)-6a, 7,8,9-tetrahydro-6Hpyrido[3,2-b]pyrrolo[1,2d][l,4]oKca3HH-4- yl)thio)pyrazin-2-yl)methanol 42 A 0 CO O—4 0, Oh oh (3S,4S)-8-(5-(((6aS,8S)-8((methoxymethoxy)methyl)6a,7,8,9-tetrahydro-6H- pyrido[3,2-b]pyrrolo[1,2d][1,4]oxazine-4-

- 9 044940

yl)thio)pyrazin-2-yl)-3-methyl2-oxa-8-azaspiro[4.5]decane-4amine 43 :/ CD Μ/\ ρ (3S,4S)-8-(5-(((6aS,8S)-8(methoxymethyl)-ba, 7,8,9tetrahydro-6H-pyrido[3,2b]pyrrolo[1,2-d] [ 1 ,4]oxazin-4yl)thio)pyrazin-2-yl)-3-methyl2-oxa-8-azaspiro[4.5]decane-4amine 44 Ο ϊ ω ¢, ο ο ρ (3S,4S)-8-(6-aMHHO-5-(((6aS,8S)8-((methoxymethoxy)methyl)6a,7,8,9-tetrahydro-6Hpyrido[3,2-b]pyrrolo[ 1,20][1,4]oxazin-4yl)thio)pyrazin-2-yl)-3-methyl2-oxa-8-azaspiro[4.5]decane-4amine 45 /^γ^ο νη ₂ CWA ^Ν > η ₂ ν* ρ (3S,4S)-8-(6-aMHHO-5-(((6aS,8S)8-(methoxymethyl)-6a, 7,8,9tetrahydro-6H-pyrido[3,2b]pyrrolo[1,2- d] [ 1,4]oxazin-4yl)thio)pyrazin-2-yl)-3 -methyl2-oxa-8-azaspiro[4.5]decane-4amine 49 Ο £ g=/ ω ^{χ ο—} C ο ο 0 (3 -((3 S,4 8)-4-amino-3 -methyl-2oxa-8-azaspiro[4.5]decan-8yl)-6-(((6a8,88)-8((methoxymethoxy)methyl) 6a,7,8,9-tetrahydro-6Hpyrido[3,2-b]pyrrolo[1,2-

- 10 044940

d] [1,4]oxazin-4-yl)thio)-5 - methylpyrazin-2-yl)methanol 50 0 1 Ν > \ JL Πχ πσ Η ₂ Π (3 -((3S,48)-4-amino-3-methyl-2oxa-8-azaspiro[4.5]decan-8yl)-6-(((6a8,8R)-8((methoxymethoxy)methyl) 6a,7,8,9-tetrahydro-6Hpyrido[3,2-b]pyrrolo[1,24][1,4]oxazin-4- yl)thio)pyrazin-2-yl)methanol 51 \/ CO ο^Λ / )=ζ V........ (3 -((3 S,4 8)-4-amino-3 -methyl-2oxa-8-azaspiro[4.5]decan-8yl)-6-(((6aK,88)-8((methoxymethoxy)methyl) 6a,7,8,9-tetrahydro-6Hpyrido[3,2-b]pyrrolo[1,24][1,4]oxazin-4yl)thio)pyrazin-2-yl)methanol 52 Ζ— ?=( / {_ζ- ° 2 J Ο ζ—/ □: / Ή ⁷ (3 -((3S,48)-4-amino-3-methyl-2oxa-8-azaspiro[4.5]decan-8yl)-6-(((6aK,8R)-8((methoxymethoxy)methyl) 6a,7,8,9-tetrahydro-6Hpyrido[3,2-b]pyrrolo[1,24][1,4]oxazin-4- yl)thio)pyrazin-2-yl)methanol 53 Η CM ¹ Ζ> ^ζ (3 -((3S,4 8)-4-amino-3-methyl-2oxa-8-azaspiro[4.5]decan-8yl)-6-((6a8,88)-8(methoxymethyl)-6a,7 ,8,9-

- 11 044940 tetrahydro-6H-pyrido[3,2b]pyrrolo[1,2-d][1,4]oxazin-4yl)pyrazin-2-yl)methanol

(3 -((3 S,4 8)-4-amino-3 -methyl-2oxa-8-azaspiro[4.5]decan-8yl)-6-((6a8,88)-8((methoxymethoxy)methyl)6a ,7,8,9-tetrahydro-6Hpyrido[3,2-b]pyrrolo[1,2d][1,4]oxazin-4-yl)pyrazin-2yl)methanol

Embodiments.

In other embodiments, the present invention includes.

1. Embodiment 1 provides a compound of formula (PV) or a pharmaceutically acceptable salt thereof:

(PV) or a pharmaceutically acceptable salt thereof, where:

where Z is -Y-M;

Y is a bond;

M is alkyl substituted with -O(alkylene)OK ^a , where R ^a is alkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl or heterocyclylalkyl; or

Z is methoxymethyloxy, methoxyethyloxy, methoxymethyl, cyclopropylmethyloxymethyl or oxetan-3-ylmethyloxymethyl; or

Z is (methoxymethoxy)methoxymethyl;

R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen, alkyl or halogen;

R ⁵ and R ⁶ are independently selected from hydrogen, alkyl or halogen;

L is a bond, O, S, S(O) or S(O) ² ;

R ⁹ is hydrogen, alkyl, halogen or amino;

R ¹⁰ is hydrogen, alkyl, halogen, hydroxy, hydroxyalkyl or -CD ₂ OH; And

R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

NH ₂

XWV* -

2. In Embodiment 2, the compound of Embodiment 1 or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyloxy, methoxyethyloxy, methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, cyclopropylmethyloxymethyl

- 12 044940 or oxetan-3-ylmethyloxymethyl.

3. In Embodiment 3, the compound of Embodiment 2, or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, cyclopropylmethyloxymethyl, or oxetan-3-ylmethyloxymethyl.

4. In embodiment 4, the compound of embodiment 3, or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, or cyclopropylmethyloxymethyl.

5. In embodiment 5, the compound of embodiment 4, or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, or methoxyethyloxymethyl.

6. In Embodiment 6, the compound of Embodiment 1, or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen and alkyl.

7. In Embodiment 7, the compound of Embodiment 1, or a pharmaceutically acceptable salt thereof, wherein one of ^R1 , ^R2 , ^R3, and ^R4 is hydrogen, and the remaining three of ^R1 , ^R2 , ^R3, and ^R4 are independently selected from hydrogen, methyl and fluorine.

8. In Embodiment 8, the compound of Embodiment 1 or a pharmaceutically acceptable salt thereof, wherein three of ^R1 , ^R2 , ^R3 and ^R4 are hydrogen and the remaining one of ^R1 , ^R2 , ^R3 and ^R4 is selected from hydrogen, methyl and fluorine.

9. In Embodiment 9, the compound of Embodiment 1, or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ² , R ³ and R ⁴ are each hydrogen.

10. In embodiment 10, the compound of embodiment 1 or a pharmaceutically acceptable salt thereof, wherein L is S, S(O) or S(O) ₂ .

11. In embodiment 11, the compound of embodiment 10, or a pharmaceutically acceptable salt thereof, wherein L is S.

12. In embodiment 12, the compound of embodiment 10, or a pharmaceutically acceptable salt thereof, wherein L is S(O) ₂ .

13. In Embodiment 13, the compound of Embodiment 1, or a pharmaceutically acceptable salt thereof, wherein R ⁹ is hydrogen, amino, or alkyl.

14. In embodiment 14, the compound of embodiment 13, or a pharmaceutically acceptable salt thereof, wherein R ⁹ is hydrogen.

15. In Embodiment 15, the compound of Embodiment 1, or a pharmaceutically acceptable salt thereof, wherein R ¹⁰ is hydroxyalkyl.

16. In embodiment 16, the compound of embodiment 15, or a pharmaceutically acceptable salt thereof, wherein R ¹⁰ is hydroxymethyl.

17. In Embodiment 17, the compound of Embodiment 1 or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyloxy, methoxyethyloxy, methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, cyclopropylmethyloxymethyl or oxetan-3-ylmethyloxymethyl;

R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen, alkyl and halogen;

L is S, S(O) or S(O) ₂ ;

R ⁹ is hydrogen, amino or alkyl; and R ¹⁰ is hydroxyalkyl.

18. In Embodiment 18, the compound of Embodiment 1 or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from /VW

19. In embodiment 18, the compound of embodiment 18, or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² , together with the carbon atom to which they are attached, form a ring selected from

20. In embodiment 20, the compound of embodiment 18, or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

- 13 044940

21. In embodiment 21, the compound of embodiment 17 or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

LLLG

22. In embodiment 22, the compound of embodiment 21, or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

23. In embodiment 23, the compound of embodiment 21, or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

24. In embodiment 24, the compound of Embodiment 1 or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyloxy, methoxyethyloxy, methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, cyclopropylmethyloxymethyl or oxetan-3-ylmethyloxymethyl;

R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen, alkyl and halogen;

L is S, S(O) or S(O) ₂ ;

R ⁹ is hydrogen, amino or alkyl;

R ¹⁰ is hydroxyalkyl; And

R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

ΛΛΛΤ

25. In Embodiment 25, the compound of Embodiment 1 or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, or methoxyethyloxymethyl;

R ¹ , R ² , R ³ and R ⁴ are hydrogen;

L is S, S(O) or S(O) ₂ ;

R ⁹ is hydrogen;

R ¹⁰ is hydroxyalkyl; And

R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

26. In embodiment 26, the compound of embodiment 25, or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

- 14044940

27. In embodiment 27, the compound of embodiment 25, or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

28. In embodiment 28, the compound of embodiment 25, or a pharmaceutically acceptable salt thereof, wherein L is S.

29. In embodiment 29, the compound of formula (IIB):

or a pharmaceutically acceptable salt thereof, wherein Z is -Y-M;

Y is a bond;

M is alkyl substituted with alkoxy or -O(αalkylene)OR ^a where R ^a is alkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl or heterocyclylalkyl;

or Z is alkoxy, cycloalkylmethyloxymethyl, heterocyclyloxy, heterocyclylmethyloxymethyl; where if Z is alkoxy, it is optionally substituted with methoxy;

R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen, alkyl or halogen;

R ⁵ and R ⁶ are independently selected from hydrogen, alkyl or halogen;

L is a bond, O, S, S(O) or S(O) ₂ ;

R ⁹ is hydrogen, alkyl, halogen or amino;

R ¹⁰ _R 11 is hydrogen, alkyl, halogen, hydroxy, hydroxyalkyl or -CD2OH; and and R ¹² together with the carbon atom to which they are attached form a ring selected from

30. In embodiment 30, the compound of embodiment 29, or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyloxy, methoxyethyloxy, methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, cyclopropylmethyloxymethyl, or oxetan-3-ylmethyloxymethyl.

31. In embodiment 31, the compound of embodiment 30, or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, cyclopropylmethyloxymethyl, or oxetan-3-ylmethyloxymethyl.

32. In embodiment 32, the compound of embodiment 31, or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, or cyclopropylmethyloxymethyl.

33. In embodiment 33, the compound of embodiment 32 or a pharmaceutical thereof

- 15 044940 highly acceptable salt, where Z is methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl or methoxyethyloxymethyl.

34. In embodiment 34, the compound of embodiment 29, or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen and alkyl.

35. In embodiment 35, the compound of embodiment 34, or a pharmaceutically acceptable salt thereof, wherein one of ^R1 , ^R2 , ^R3, and ^R4 is hydrogen, and the remaining three of ^R1 , ^R2 , ^R3, and ^R4 are independently selected from hydrogen, methyl and fluorine.

36. In embodiment 36, the compound of embodiment 34 or a pharmaceutically acceptable salt thereof, wherein three of ^R1 , ^R2 , ^R3 and ^R4 are hydrogen and the remaining one of ^R1 , ^R2 , ^R3 and ^R4 is selected from hydrogen, methyl and fluorine.

37. In embodiment 37, the compound of embodiment 34, or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ² , R ³ and R ⁴ are each hydrogen.

38. In embodiment 38, the compound of embodiment 29, or a pharmaceutically acceptable salt thereof, wherein L is S, S(O) or S(O) ₂ .

39. In embodiment 39, the compound of embodiment 38, or a pharmaceutically acceptable salt thereof, wherein L is S.

40. In embodiment 49, the compound of embodiment 38, or a pharmaceutically acceptable salt thereof, wherein L is S(O) ₂ .

41. In embodiment 41, the compound of embodiment 29, or a pharmaceutically acceptable salt thereof, wherein R ⁹ is hydrogen, amino, or alkyl.

42. In embodiment 42, the compound of embodiment 41, or a pharmaceutically acceptable salt thereof, wherein R ⁹ is hydrogen.

43. In embodiment 43, the compound of embodiment 29, or a pharmaceutically acceptable salt thereof, wherein R ¹⁰ is hydroxyalkyl.

44. In embodiment 44, the compound of embodiment 43, or a pharmaceutically acceptable salt thereof, wherein R ¹⁰ is hydroxymethyl.

45. In embodiment 45, the compound of embodiment 29, or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyloxy, methoxyethyloxy, methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, cyclopropylmethyloxymethyl, or oxetan-3-ylmethyloxymethyl;

R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen, alkyl or halogen;

L is S, S(O) or S(O) ₂ ;

R ⁹ is hydrogen, amino or alkyl; And

R ¹⁰ is hydroxyalkyl.

46. In embodiment 46, the compound of embodiment 29, or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

47. In embodiment 47, the compound of embodiment 46, or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with a ring selected from the carbon atom to which they are attached form

48. In embodiment 48, the compound of embodiment 46, or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

49. In embodiment 49, the compound of embodiment 45, or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

- 16 044940

50. In Embodiment 50, the compound of Embodiment 49 or a pharmaceutical thereof, together with the carbon atom to which they are attached, form a skilfully acceptable salt, wherein R ¹¹ and R ¹² are a ring selected from

51. In embodiment 51, the compound of embodiment 49, or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

52. In embodiment 52, the compound of embodiment 29 or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyloxy, methoxyethyloxy, methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, cyclopropylmethyloxymethyl or oxetan-3-ylmethyloxymethyl;

R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen, alkyl or halogen;

L is S, S(O) or S(O) ₂ ;

R ⁹ is hydrogen, amino or alkyl;

R ¹⁰ is hydroxyalkyl; And

R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

_NH2 . _nh2 . nh ₂

53. In embodiment 53, the compound of embodiment 29, or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, or methoxyethyloxymethyl;

R ¹ , R ² , R ³ and R ⁴ are hydrogen;

L is S, S(O) or S(O) ₂ ;

R ⁹ is hydrogen;

R ¹⁰ is hydroxyalkyl; And

R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

54. In embodiment 54, the compound of embodiment 53, or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

55. In embodiment 55, the compound of embodiment 53, or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from

- 17 044940

56. In embodiment 56, the compound of embodiment 53, or a pharmaceutically acceptable salt thereof, wherein L is S.

57. In embodiment 57, the compound is selected from the group consisting of:

- 18 044940

or a pharmaceutically acceptable salt thereof.

58. In embodiment 58, the connection is as in embodiment 57, where the connection is

59. In embodiment 59, the connection of embodiment 57, where the connection is

60. In embodiment 60, the connection of embodiment 57, where the connection is

61. In embodiment 61, the connection of embodiment 57, where the connection is

- 19 044940

62. In embodiment 62, the connection of embodiment 57, where the connection is

63. In embodiment 63, the connection of embodiment 57, where the connection is

64. In embodiment 64, the connection of embodiment 57, where the connection is

65. In embodiment 65, the connection of embodiment 57, where the connection is

66. In embodiment 66, the connection of embodiment 57, where the connection is

67. In embodiment 67, the connection of embodiment 57, where the connection is

- 20 044940

68. In embodiment 68 is the connection of embodiment 57, where the connection is

69. In embodiment 69, the compound of embodiment 57, wherein the compound is a pharmaceutically acceptable salt of a compound that is

70. In embodiment 70, the compound of embodiment 57, wherein the compound is a pharmaceutically acceptable salt of a compound that is

71. In embodiment 71, the compound of embodiment 57, wherein the compound is a pharmaceutically acceptable salt of a compound that is

72. In embodiment 72, the compound of embodiment 57, wherein the compound is a pharmaceutically acceptable salt of a compound that is

73. In embodiment 73, the compound of embodiment 57, wherein the compound is a pharmaceutically acceptable salt of a compound that is

- 21 044940

74. In embodiment 74, the compound of embodiment 57, wherein the compound is a pharmaceutically acceptable salt of a compound that is

75. In embodiment 75, the compound of embodiment 57, wherein the compound is a pharmaceutically acceptable salt of a compound that is

76. In embodiment 76, the compound of embodiment 57, wherein the compound is a pharmaceutically acceptable salt of a compound that is

77. In embodiment 77, the compound of embodiment 57, wherein the compound is a pharmaceutically acceptable salt of a compound that is

78. In embodiment 78, the compound of embodiment 57, wherein the compound is a pharmaceutically acceptable salt of a compound that is

79. In embodiment 79, the compound of embodiment 57, wherein the compound is a pharmaceutically acceptable salt of a compound that is

80. A pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1-79 and a pharmaceutically acceptable excipient.

81. Use of the compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1-79 for the treatment of a disease treatable by inhibition of SHP2.

82. Use according to embodiment 81 wherein the disease is cancer.

83. Use according to embodiment 82, wherein the cancer is selected from lung, stomach, liver, colon

- 22 044940 intestine, kidney, breast, pancreatitis, juvenile myelomonocytic leukemia, neuroblastoma, melanoma and acute myeloid leukemia.

General synthesis scheme.

Compounds of this description can be prepared by the methods depicted in the reaction schemes shown below.

The starting materials and reagents used in the preparation of these compounds can be purchased from commercial suppliers such as Aldrich Chemical Co., (Milwaukee, Wis.), Bachem (Torrance, Calif.), or Sigma (St. Louis, Mo.) or obtained by methods known to those skilled in the art according to the procedures set forth in the references below, such as Fieser and Fieser's Reagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd's Chemistry of Carbon Compounds, Volumes 1-5 and Supplemental (Elsevier Science Publishers, 1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991), March's Advanced Organic Chemistry, (John Wiley and Sons, 4th Edition) and Larock's Comprehensive Organic Transformations (VCH Publishers Inc., 1989). These diagrams are merely illustrative of some of the ways in which the compounds of this disclosure may be synthesized, and various modifications may be made to these schemes as will be appreciated by one skilled in the art reading this disclosure. The starting materials and intermediates and final reaction products can be isolated and purified, if desired, using conventional techniques including, but not limited to, filtration, distillation, crystallization, chromatography and the like. Such materials can be characterized using conventional means, including physical constants and spectral data.

Unless otherwise specified, the reactions described herein take place at atmospheric pressure over a temperature range of about -78°C to about 150°C, such as from about 0°C to about 125°C, and then, for example, about at room (or ambient) temperature, for example about 20°C.

General synthesis scheme.

Compounds of formula (IIB) in which L is S and other groups as defined in the Summary of the Invention can be prepared as shown and described in Scheme 1 below.

Scheme 1.

A combination of a compound of formula 1-a, where X ¹ is a halogen, where Z, R ¹ , R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are as defined in the Summary of the invention, or its predecessor, and E and A such as those defined in the Summary of the Invention, with a compound of formula 1-d, wherein M ¹ is a metal such as sodium or potassium, in the presence of a transition metal catalyst such as Pd ₂ (dba) ₃ and xanthos, under standard conditions combination gives compounds of formula (IIB). If a compound of formula 1-a is substituted with an amino group, the amino group may be protected with a suitable protecting group such as tert-butylcarbamate prior to the coupling reaction. Removal of the amino protecting group by methods well known in the art then gives the compound of formula (IIB).

Alternatively, compounds of formula (IIB) can be prepared by reacting a compound of formula 1-a with a 3-mercaptopropanoate ester in the presence of a transition metal catalyst such as Pd ₂ (dba) ₃ and xanthos under standard coupling conditions to obtain a compound of formula 1-b, where R' is an alkyl group such as 3-methylheptane. Treatment of a compound of formula 1-b with a base such as potassium t-butoxide, sodium t-butoxide, sodium methoxide and the like gives the compound of formula 1-c as a thiosalt, where M ⁺ is a metal ion such as potassium or sodium. The combination of 1-c with a compound of formula 1-e or formula 1-f, where X ² or X ³ is a halogen, gives a compound of formula (IIB), where Z 1 is a group of formula (a) or (b), respectively.

- 23 044940

Compounds of formula 1-a, where X ¹ is a halogen, E is O and A, E, Z, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ such as defined in the Summary of the invention, or their precursors can be prepared as shown and described in Methods 1 and 2 below.

Method (1).

Combining a compound of formula 2-b, wherein R ⁵ and R ⁶ are as defined in the Summary of the Invention, or its precursor with an alcohol of formula 2-a, where PG is a suitable amino protecting group such as Boc, under Mitsunobu conditions, e.g. , using diethyl azodicarboxylate and triphenylphosphine, gives the compound of formula 2-c. The compounds of formula 2-b are commercially available or can be prepared by methods well known in the art. For example, 2-fluoropyridin-3-ol is commercially available. The compounds of formula 2-a are commercially available or they can be prepared by methods well known in the art. For example, tert-butyl (S)-2-(hydroxymethyl)pyrrolidine-1-carboxylate, tert-butyl (R)-2(hydroxymethyl)pyrrolidine-1-carboxylate, tert-butyl (2S,4S)-4-fluoro- 2-(hydroxymethyl)-pyrrolidine-1-carboxylate, tert-butyl (2S,4R)-4-fluoro-2-(hydroxymethyl)pyrrolidine-1-carboxylate, tert-butyl (S)4,4-difluoro-2-(hydroxymethyl )pyrrolidine-1-carboxylate, tert-butyl (2S,4R)-2-(hydroxymethyl)-4methoxypyrrolidine-1-carboxylate, tert-butyl (2S,4S)-2-(hydroxymethyl)-4-methoxypyrrolidine-1-carboxylate, tert -butyl (S)-6-(hydroxymethyl)-5-azaspiro[2.4]heptane-5-carboxylate, tert-butyl (R)-3(hydroxymethyl)morpholine-4-carboxylate, tert-butyl (S)-3- (hydroxymethyl)morpholine-4-carboxylate, tert-butyl (R)-2-(hydroxymethyl)azetidine-1-carboxylate, tert-butyl (S)-2-(hydroxymethyl)azetidine-1-carboxylate are commercially available.

Removal of the amino protecting group gives the compound of formula 2-d. For example, the Boc group can be cleaved under acidic conditions such as HCl in dioxane. Cyclization of compound 2-d with a base such as K ₂ CO ₃ , sodium carbonate and the like gives the compound of formula 2-d. Quotation of compound 2 using an alkyl lithium such as n-BuLi followed by capture with iodine gives compound of formula 1-a.

Method (2).

- 24 044940

Alternatively, a combination of a compound of formula 3-a, where X ¹ is a halogen, R ⁵ and R ⁶ as defined in the Summary of the invention, and an alcohol of formula 2-a, where PG is an amino protecting group such as Boc, under Mitsunobu conditions , for example, using diethyl azodicarboxylate and triphenylphosphine, gives the compound of formula 3-b. The compounds of formula 3-a are commercially available or they can be prepared by methods well known in the art. For example, 2-fluoro-4-iodopyridin-3-ol is commercially available. Compound 3-c is converted to compound of formula 1-a as described in Scheme 1 above.

Application.

Src homology domain phosphatase 2 (SHP2) is a protein tyrosine phosphatase encoded by the PTPN1 gene 1 that is involved in many cellular functions, including proliferation, differentiation, cell cycle maintenance, and migration. SHP2 is involved in signaling through the Ras mitogen-activated protein kinase, JAK-STAT, or phosphoinositol 3-kinase-AKT pathway. SHP2 mediates the activation of the MAP kinases Erk1 and Erk2 (Erk1/2, Erk) by receptor tyrosine kinases such as ErbB1, ErbB2 and c-Met.

SHP2 has two N-terminal Src homology 2 domains (N-SH2 and C-SH2), a catalytic domain (PTP), and a C-terminal tail. Two SH2 domains control the subcellular localization and functional regulation of SHP2. The molecule exists in an inactive conformation, suppressing its activity through a binding network including residues from the N-SH2 and PTP domains. In response to growth factor stimulation, SHP2 binds to specific tyrosine-phosphorylated sites on docking proteins such as Gab1 and Gab2 through its SH2 domains. This causes a conformational change that leads to SHP2 activation.

Mutations in PTPN11 have been identified in several human diseases, such as Noonan syndrome, Leopard syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, acute myeloid leukemia, and breast, lung, and colon cancers. SHP2 is an important downstream signaling molecule for various receptor tyrosine kinases, including platelet-derived growth factor receptors (PDGF-R), fibroblast growth factor receptors (FGF-R), and epidermal growth factor receptors (EGFR). SFLP2 is also an important downstream signaling molecule for activation of the mitogen-activated protein (MAP) kinase pathway, which can lead to cell transformation, a prerequisite for cancer development. Knockdown of SFIP2 significantly suppresses the growth of lung cancer cell lines with SFLP2 mutation or EML4/ALK translocations, as well as EGFR amplified breast cancer and esophageal cancer. SFIP2 is also upregulated downstream of oncogenes in gastric carcinoma, anaplastic large cell lymphoma, and glioblastoma.

Noonan syndrome (NS) and Leopard syndrome (LS): PTPN11 mutations cause LS (multiple lentiginosis, electrocardiographic conduction abnormalities, familial hypertelorism, pulmonary stenosis, abnormal genitalia, growth retardation, sensorineural deafness) and NS (congenital anomalies including heart defects, craniofacial anomalies and short stature). Both disorders are part of a family of autosomal dominant syndromes caused by germline mutations in components of the RAS/RAF/MEK/ERK mitogen-activating protein kinase pathway required for normal cell growth and differentiation. Aberrant regulation of this pathway has profound effects, particularly on cardiac development, leading to various abnormalities including valvuloseptal defects and/or hypertrophic cardiomyopathy (HCM). Disturbances in the MAPK signaling pathway have been identified as central to these disorders, and several candidate genes in this pathway have been identified in humans, including mutations in KRAS, NRAS, SOS1, RAF1, BRAF, MEK1, MEK2, SHOC2, and CBL. The gene most frequently mutated in NS and LS is PTPN1 1. Germline mutations in PTPN1 1 (SFIP2) are found in -50% of cases with NS and in almost all patients with LS, which shares features with NS. For NS, the Y62D and Y63C substitutions in the protein are largely invariant and are among the most common mutations. Both of these mutations affect the catalytically inactive conformation of SFIP2 without disrupting the binding of the phosphatase to its phosphorylated signaling partners.

Juvenile myelomonocytic leukemia (JMML): - Somatic mutations in PTPN1 1 (SFIP2) occur in approximately 35% of patients with JMML, a childhood myeloproliferative disorder (MPD). These gain-of-function mutations are usually point mutations in the N-SH2 domain or in the phosphatase domain that prevent the self-inhibition between the catalytic domain and the N-SH2 domain, resulting in SFIP2 activity.

Acute myeloid leukemia: PTPN1 1 mutations have been identified in: -10% of childhood acute leukemias such as myelodysplastic syndrome (MDS); -7% B-cell acute lymph regional leukemia (B-ALL); and -4% acute myeloid leukemia (AML).

NS and leukemia mutations cause changes in amino acids located at the interface formed by the N-SH2 and PTP domains in the self-inhibitory conformation of SHP2, disrupting the inhibitory intramolecular interaction, resulting in hyperactivity of the catalytic domain.

SHP2 acts as a positive regulator in receptor tyrosine kinase (RTK) signaling. Cancers containing RTK changes (EGFR amp, Her2 amp, FGFR amp, Met 31 15, translocated/activated RTK, i.e. ALK, BCR/ABL) include cancers of the esophagus, breast, lung, colon, stomach , glioma, head and neck.

Esophageal cancer (or esophageal cancer) is a malignant neoplasm of the esophagus. There are various subtypes, most notably squamous cell carcinoma (<50%) and adenocarcinoma. RTK expression levels are high in esophageal adenocarcinoma and squamous cell carcinoma. Therefore, the SHP2 inhibitor of the present invention can be used for innovative treatment strategies.

Breast cancer is a major type of cancer and a leading cause of death in women when patients develop resistance to current drugs. There are four main subtypes of breast cancer, including luminal A, luminal B, Her2-like, and triple-negative/basal. Triple-negative breast cancer (TNBC) is an aggressive breast cancer that does not have specific targeted therapy. Epidermal growth factor receptor I (EGFR) has emerged as a promising target for TNBC. Inhibition of Her2 as well as EGFR via SFIP2 may be a promising treatment for breast cancer.

Lung cancer - NSCLC is currently the leading cause of cancer mortality, accounting for approximately 85% of lung cancers (primarily adenocarcinomas and squamous cell carcinomas). Although cytotoxic chemotherapy remains an important part of treatment, tumors are more likely to benefit from targeted therapy based on genetic changes such as EGFR and ALK.

Colon Cancer - Approximately 30% to 50% of colorectal tumors are known to have mutated (abnormal) KRAS, and BRAF mutations occur in 10-15% of colorectal cancers. For the subset of patients whose colorectal tumors have demonstrated EGFR overexpression, these patients demonstrate a favorable clinical response to anti-EGFR therapy.

Stomach cancer is one of the most common types of cancer. Aberrant expression of tyrosine kinases, which is reflected in aberrant tyrosine phosphorylation in gastric cancer cells, is known in the art. Three receptor tyrosine kinases, c-met (HGF receptor), FGF receptor 2, and erbB2/neu, are frequently amplified in gastric carcinomas. Thus, disruption of different signaling pathways may contribute to the progression of different types of gastric cancer.

Neuroblastoma is a pediatric tumor of the developing sympathetic nervous system, accounting for approximately 8% of all childhood cancers. Genomic alterations in the anaplastic lymphoma kinase (ALK) gene are believed to contribute to the pathogenesis of neuroblastoma.

Squamous cell carcinoma of the head and neck (SCCHN). High levels of EGFR expression correlate with poor prognosis and resistance to radiation therapy in various forms of cancer, mainly squamous cell carcinoma of the head and neck (SCCHN). Blocking EGFR signaling results in inhibition of receptor stimulation, cell proliferation, and reduced invasiveness and metastasis. Therefore, EGFR is a major target for novel antitumor therapy in SCCFIN.

The present invention relates to compounds capable of inhibiting SHP2 activity. The invention also provides a method for preparing the compounds of the invention and pharmaceutical preparations containing such compounds. Another aspect of the present invention relates to a method of treating SHP2-mediated disorders, comprising the step of administering to a patient in need thereof a therapeutically effective amount of a compound of Formula IIB, as defined in the Summary of the Invention.

In some embodiments, the present invention relates to the above method, wherein said SHP2-mediated disorders are cancers selected from, but not limited to: JMML; AML; MDS; B-ALL; neuroblastoma; esophageal carcinoma; breast cancer; lungs' cancer; colon cancer; stomach cancer, head and neck cancer.

The compounds of the present invention may also be used in the treatment of other diseases or conditions associated with aberrant SFIP2 activity. Thus, in another aspect, the invention relates to a method of treating a disorder selected from: JMML; AML; MDS; B-ALL; neuroblastoma; esophageal cancer; breast cancer; lung cancer; colon cancer; stomach cancer, head and neck cancer.

The SFIP2 inhibitor of the present invention can be effectively combined with another pharmacologically active compound or two or more other pharmacologically active compounds, especially in the treatment of cancer. For example, a compound of the present invention, or a pharmaceutically acceptable salt thereof, as defined above, can be administered simultaneously, sequentially, or separately in combination with one or more agents selected from chemotherapeutic agents, for example, mitotic inhibitors such as taxane, vinca alkaloid, paclitaxel, docetaxel, vincristine, vinblastine, vinorelbine or vinflunine and other anticancer agents such as cisplatin, 5-fluorouracil or 5-fluoro-2-4-(III,3H)pyrimidinedione (5FU), flutamide or gemcitabine. Such combinations may have significant therapeutic benefits, including synergistic activity.

In some embodiments, the present invention relates to the above method, wherein said compound is administered parenterally.

- 26 044940

In some embodiments, the present invention relates to the above method, wherein the compound is administered intramuscularly, intravenously, subcutaneously, orally, pulmonaryly, intrathecally, topically or intranasally.

In some embodiments, the present invention relates to the above method, wherein said compound is administered systemically.

In some embodiments, the present invention relates to the above method, wherein said patient is a mammal.

In some embodiments, the present invention relates to the above method, wherein said patient is a primate.

In some embodiments, the present invention relates to the above method, wherein said patient is a human.

In another aspect, the present invention relates to a method of treating an SHP2-mediated disorder, comprising the step of: administering to a patient in need thereof a therapeutically effective amount of a chemotherapeutic agent in combination with a therapeutically effective amount of a compound of Formula I, as defined in the Summary of the Invention.

In addition to human cancer, SHP2 inhibition also has therapeutic potential for the treatment of systemic lupus erythematosus, rheumatoid arthritis, and fibrosis.

Testing.

The SHP2 inhibitory effect of compounds of formula (IIB) can be tested using the in vitro assay described in Biological Examples 1 below.

Pharmaceutical compositions.

In general, the compounds of this disclosure are administered in a therapeutically effective amount by any of the acceptable routes of administration for agents that serve similar purposes. Therapeutically effective amounts of the compounds of the present disclosure may range from about 0.01 to about 500 mg per kg of patient body weight per day, which can be administered in single or multiple doses. A suitable dosage level may be from about 0.1 to about 250 mg/kg per day; from about 0.5 to about 100 mg/kg per day. A suitable dosage level may be from about 0.01 to about 250 mg/kg per day, from about 0.05 to about 100 mg/kg per day, or from about 0.1 to about 50 mg/kg per day. Within this range, the dose may be from about 0.05 to about 0.5, from about 0.5 to about 5, or from about 5 to about 50 mg/kg per day. For oral administration, the compositions may be presented in the form of tablets containing from about 1.0 to about 1000 milligrams of active ingredient, in particular about 1, 5, 10, 15, 20, 25, 50, 75, 100, 150, 200 , 250, 300, 400, 500, 600, 750, 800, 900 and 1000 milligrams of the active ingredient. The actual amount of the compound as described herein, i.e. active ingredient will depend on many factors, such as the severity of the disease being treated, the age and relative health of the patient, the effectiveness of the compound used, the route and form of administration and other factors.

In general, the compounds of the present disclosure are administered as pharmaceutical compositions by any of the following routes: oral, systemic (eg, transdermal, intranasal, or suppository), or parenteral (eg, intramuscular, intravenous, or subcutaneous) administration. The preferred route of administration is oral, using a convenient daily dosage schedule that can be adjusted depending on the extent of the lesion. The compositions may take the form of tablets, pills, capsules, semi-solids, powders, sustained release formulations, solutions, suspensions, elixirs, aerosols or any other suitable compositions.

The choice of formulation depends on various factors, such as the route of administration of the drug (eg, for oral administration, formulations in the form of tablets, pills or capsules, including enteric-coated or sustained-release tablets, pills or capsules, are preferred) and the bioavailability of the drug substance.

The compositions generally consist of a compound as described herein in combination with at least one pharmaceutically acceptable excipient. Suitable excipients are non-toxic, facilitate administration, and do not adversely affect the therapeutic effect of the compound herein. Such excipient may be any solid, liquid, semi-solid or, in the case of an aerosol composition, gaseous excipient that is generally available to one skilled in the art.

Solid pharmaceutical excipients include starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, skim milk powder and the like. Liquid and semi-solid excipients may be selected from glycerin, propylene glycol, water, ethanol and various oils, including petroleum, animal, vegetable or synthetic oils, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. Preferred liquid carriers, especially for injection solutions, include water, saline, aqueous dextrose and glycols.

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The compounds can be formulated for parenteral administration by injection, for example, by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, for example, in ampoules or in multi-dose containers with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain auxiliary substances such as suspending, stabilizing and/or dispersing agents. The formulations may be presented in single-dose or multi-dose containers, such as sealed ampoules and vials, and may be stored in powder form or under freeze-dried (lyophilized) conditions, requiring only the addition of a sterile liquid carrier, such as saline or sterile pyrogen-free water, immediately prior to use. Injection solutions and suspensions prepared for immediate administration can be prepared from the sterile powders, granules and tablets described previously.

Formulations for parenteral administration include aqueous and non-aqueous (oily) sterile injectable solutions of the active compounds, which may contain antioxidants, buffers, bacteriostatic agents and solutes that render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions, which may include suspending agents and thickening agents. Suitable lipophilic solvents or carriers include fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol or dextran. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the compounds, allowing the preparation of highly concentrated solutions.

In addition to the formulations described previously, the compounds may also be formulated as depot formulations. Such long-acting formulations may be administered by implantation (eg, subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds may be formulated with suitable polymeric or hydrophobic materials (eg, as an emulsion in a suitable oil) or ion exchange resins, or as sparingly soluble derivatives, eg, as a sparingly soluble salt.

For buccal or sublingual administration, the compositions may take the form of tablets, lozenges, lozenges or gels prepared in conventional manner. Such compositions may contain the active ingredient in a flavored base such as sucrose and gum acacia or tragacanth.

The compounds can also be formulated as rectal compositions such as suppositories or retention enemas, for example containing conventional suppository bases such as cocoa butter, polyethylene glycol or other glycerides.

Certain compounds disclosed herein may be administered locally, that is, not by systemic administration. This involves applying the compound described herein externally to the epidermis or oral cavity, and instilling such compound into the ear, eye and nose so that the compound does not significantly enter the bloodstream. In contrast, systemic administration refers to oral, intravenous, intraperitoneal and intramuscular administration.

Formulations suitable for topical application include liquid or semi-liquid preparations suitable for penetration through the skin to the site of inflammation, such as gels, liniments, lotions, creams, ointments or pastes and drops suitable for administration to the eyes, ear or nose. The topical active ingredient may contain, for example, from 0.001% to 10% w/w (by weight) of the composition. In some embodiments, the active ingredient may be no more than 10% w/w. In other embodiments, it may be less than 5% w/w. In some embodiments, the active ingredient may range from 2% w/w. up to 5% w/w In other embodiments, it may range from 0.1% to 1% w/w. composition.

For administration by inhalation, the compounds may be conveniently delivered from an insufflator, a pressurized aerosol can, or other convenient aerosol spray delivery means. Aerosol packages may contain a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the standard dosage unit may be determined by the installation of a valve to deliver the metered amount. Alternatively, for administration by inhalation or insufflation, the compounds of the present disclosure may take the form of a dry powder composition, for example a powder mixture of the compound and a suitable powder base such as lactose or starch. The powder composition may be presented in unit dosage form in, for example, capsules, cartridges, gelatin or blister packs, from which the powder may be administered by means of an inhaler or insufflator. Other suitable pharmaceutical excipients and their compositions are described in Remington's Pharmaceutical Sciences, edited by E. W. Martin (Mack Publishing Company, 20th ed., 2000).

The level of compound in the formulation may vary within the full range used by those skilled in the art. Typically the composition will contain, in percentage by weight (wt.%) of

- 28 044940 about 0.01-99.99 wt.% of the compound according to the present description based on the entire composition, and the balance is one or more suitable pharmaceutical excipients. For example, the compound is present at a level of about 1-80 wt.%.

Combinations and combination therapies.

The compounds described herein may be used in combination with one or more other drugs in the treatment of diseases or conditions for which the compounds described herein or other drugs may be of use. Such other drug(s) may be administered in a manner and in an amount typically administered concurrently or sequentially with a compound of formula (IIB). When a compound of the present disclosure is used concomitantly with one or more other drugs, a pharmaceutical composition in unit dosage form containing such other drugs and the compound of formula (IIB) is preferred. However, combination therapy may also include therapies in which a compound of the present description and one or more other drugs are administered in different overlapping schedules. It is also contemplated that when used in combination with one or more other active ingredients, the compounds of formula (IIB) and the other active ingredients can be used in lower doses than when used alone.

Therefore, pharmaceutical compositions of formula (IIB) also include those that contain one or more other drugs in addition to the compound of formula (IIB).

The above-mentioned combinations include combinations of a compound herein not only with one other drug, but also with two or more other active drugs. Likewise, a compound disclosed herein may be used in combination with other drugs that are used to prevent, treat, control, alleviate, or reduce the risk of diseases or conditions that benefit from a compound disclosed herein. Such other drugs may be administered in the manner and in the amount usually used, simultaneously or sequentially with the compound of formula (IIB). When a compound described herein is used concomitantly with one or more other drugs, a pharmaceutical composition containing such other drugs in addition to the compound described herein may be used. Accordingly, pharmaceutical compositions of formula (IIB) also include those that also contain one or more other active ingredients in addition to the compound described herein. The weight ratio of the compound described herein to the second active ingredient may vary and will depend on the effective dose of each ingredient. Typically an effective dose of each will be used.

If a subject in need suffers from, or is at risk of developing, cancer, the subject may be treated with a compound herein in any combination with one or more other anti-cancer agents, including, but not limited to:

MAP kinase pathway inhibitors (RAS/RAF/MEK/ERK), including but not limited to: vemurafanib (PLX4032), dabrafenib, encorafenib (LGX818), TQ-B3233, XL-518 (Cas No. 1029872-29-4, available from ACC Corp); trametinib, selumatinib (AZD6244), TQ-B3234, PD184352, PD325901, TAK733, pimasertinib, binimetinib, refametinib, cobimetinib (GDC-0973), AZD8330, BVD-523, LTT462, ulixertinib, AMG510, ARS853 and any RAS inhibitors described in patents WO 2016049565, WO 2016164675, WO 2016168540, WO 2017015562, WO 2017058728, WO 2017058768, WO 2017058792, WO 2017058805, WO 20170 58807, WO 2017058902, WO 2017058915, WO 2017070256, WO 2017087528, WO 2017100546, WO 2017172979, WO 2017201161, WO 2018064510, WO 2018068017, WO 2018119183.

CSF1R inhibitors (PLX3397, LY3022855, etc.) and CSF1R antibodies (IMC-054, RG7155) TGF receptor kinase beta inhibitor such as LY2157299.

a BTK inhibitor such as ibrutinib; BCR-ABL inhibitors: imatinib (Gleevec®); inilotinib hydrochloride; nilotinib (Tasigna®); dasatinib (BMS-345825); bosutinib (SKI-606); ponatinib (AP24534); bafetinib (INNO406); danusertib (PHA-739358), AT9283 (CAS 1133385-83-7); saracatinib (AZD0530); and A/-[2-[(15,4R)-6-[[4-(cyclobutylamino)-5-(trifluoromethyl)-2-pyrimidinyl]amino]-1,2,3,4tetrahydronaphthalene-1,4-imine -9-yl]-2-oxoethyl]acetamide (PF-03814735, CAS 942487-16-3).

ALK inhibitors: PF-2341066 (XALKOPJ®; crizotinib); 5-chloro-N4-(2-(isopropylsulfonyl)phenyl)N2-(2-methoxy-4-(4-(4-methylpiperazin-1 -yl)piperidin-1 -yl)phenyl)pyrimidine-2,4-diamine ; GSK1838705 A; CH5424802; ceritinib (ZYKADIA); TQ-B3139, TQ-B3101 PI3K inhibitors: 4-[2-(1H-indazol-4-yl)-6-[[4(methylsulfonyl)piperazin-1-yl]methyl]thieno[3,2-c1]pyrimidine -4-yl]morpholine (also known as GDC 0941 and described in PCT Publication Nos. WO 09/036082 and WO 09/055730), 2-methyl-2-[4-[3-methyl-2-oxo-8( quinolin-3-yl)-2,3-dihydroimidazo[4,5-c]quinolin-1-yl]phenyl]propionitrile (also known as BEZ 235 or NVP-BEZ 235 and described in PCT Publication No. WO 06/122806) .

Vascular epithelial growth factor (VEGF) receptor inhibitors: bevacizumab (sold under the trade name Avastin® from Genentech/Roche), axitinib, (N-methyl-2-[[3-[(E)-2-niridine-29 044940

2-ylethenyl]-1H-indazol-6-yl]sulfanyl]benzamide, also known as AGO13736 and described in PCT Publication No. WO 01/002369), brivanib alaninate ((S)-((R)-1-(4- (4-fluoro-2-methyl-1H-indol-5-yloxy)-5methylpyrrolo[2,1-f][1,2,4]triazin-6-yloxy)propan-2-yl)2-aminopropanoate, also known as BMS582664), motesanib (M-(2,3-dihydro-3,3-dimethyl-1H-indol-6-yl)-2-[(4-pyridinylmethyl)amino]-3 pyridinecarboxamide and described in the PCT publication No. WO 02/066470), paziretotide (also known as SOM230 and described in PCT Publication No. WO 02/010192), sorafenib (marketed under the trade name Nexavar®); AL-2846 MET inhibitor such as foretinib, carbozantinib, or crizotinib.

FLT3 inhibitors - sunitinib malate (marketed under the trade name Sutent® from Pfizer); PKC412 (midostaurin); tanutinib, sorafenib, lestaurtinib, KW-2449, quizartinib (AC220) and crenolanib.

Epidermal growth factor inhibitors (EGFR): gefitnib (sold under the trade name Iressa®), N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[[(3S)-tetraαhydro-3-furanyl] hydroxy]-6-quinazolinyl]4(dimethylamino)-2-butenamide, sold under the trade name Tovok® from Boehringer Ingelheim), cetuximab (marketed under the trade name Erbitux® from Bristol-Myers Squibb), panitumumab (marketed under the trade name Vectibix® from Amgen).

HER2 receptor inhibitors: trastuzumab (sold under the trade name Herceptin® from Genentech/Roche), neratinib (also known as HKI-272, (2E)-N-[4-[[3-chloro-4-[(pyridin-2yl) methoxy]phenyl]amino]-3-cyano-7-ethoxyquinolin-6-yl]-4-(dimethylamino)but-2-enamide and described in PCT Publication No. WO 05/028443), lapatinib or lapatinib ditosylate (sold under the trade name Tykerb® from GlaxoSmithKline); trastuzumab emtansine (in the US ado-trastuzumab emtansine, trade name Kadcyla) is an antibody-drug conjugate consisting of the monoclonal antibody trastuzumab (Herceptin) bound to the cytotoxic agent mertansine (DM1);

HER dimerization inhibitors: pertuzumab (sold under the trade name Omnitarg®, from Genentech).

CD20 antibodies: rituximab (marketed under the trade names Riuxan® and MabThera® from Genentech/Roche), tositumomab (marketed under the trade names Bexxar® from GlaxoSmithKline), ofatumumab (marketed under the trade name Arzerra® from GlaxoSmithKline).

Tyrosine kinase inhibitors: erlotinib hydrochloride (sold under the trade name Tarceva® from Genentech/Roche), linifanib (N-[4-(3-amino-1H-indazol-4-yl)phenyl]-N'-(2-fluoro-5 - methylphenyl)urea, also known as ABT 869, available from Genentech), sunitinib malate (marketed under the trade name Sutent® from Pfizer), bosutinib (4-[(2,4-dichloro-5-methoxyphenyl)amino]-6methoxy- 7-[3-(4-methylpiperazin-1-yl)propoxy]quinoline-3-carbonitrile, also known as SKI-606 and described in US Patent No. 6,780,996), dasatinib (marketed under the trade name Sprycel® from Bristol-Myers Squibb ), armala (also known as pazopanib, sold under the trade name Votrient® from GlaxoSmithKline), imatinib, and imatinib mesylate (marketed under the trade names Gilvec® and Gleevec® from Novartis).

DNA synthesis inhibitors: capecitabine (sold under the trade name Xeloda® from Roche), gemcitabine hydrochloride (sold under the trade name Gemzar® from Eli Lilly and Company), nelarabine ((2R3S,4R,5R)-2-(2-amino-6 -methoxypurin-9-yl)-5-(hydroxymethyl)oxolane-3,4diol, sold under the trade names Arranon® and Atriance® from GlaxoSmithKline).

Antineoplastic agents: Oxaliplatin (marketed under the trade name Eloxatin® from Sanofi-Aventis and described in US Pat. No. 4,169,846).

Human granulocyte colony-stimulating factor (G-CSF) modulators: filgrastim (marketed under the trade name Neupogen® from Amgen).

Immunomodulators: afutuzumab (available from Roche®), pegfilgrastim (sold under the trade name Neulasta® from Amgen), lenalidomide (also known as CC-5013, sold under the trade name Revlimid®), thalidomide (sold under the trade name Thalomid®);

CD40 inhibitors: dacetuzumab (also known as SGN-40 or huS2C6, available from Seattle Genetics, Inc); Pro-apoptotic receptor agonists (PARA): dulanermine (also known as AMG-951, available from Amgen/Genentech).

Hedgehog antagonists: 2-chloro-N-[4-chloro-3-(2-pyridinyl)phenyl]-4-(methylsulfonyl)benzamide (also known as GDC-0449 and described in PCT Publication No. WO 06/028958);

Phospholipase A2 inhibitors: anagrelide (sold under the trade name Agrylin®);

BCL-2 inhibitors: 4-[4-[[2-(4-chlorophenyl)-5,5-dimethyl-1-cyclohexen-1-yl]methyl]-1piperazinyl]-N-[[4-[[(1R )-3-(4-morpholinyl)-1-[(phenylthio)methyl]propyl]amino]-3[(trifluoromethyl)sulfonyl]phenyl]sulfonyl]benzamide (also known as ABT-263 and described in PCT Publication No. WO 09 /155386);

MCl-1 inhibitors: MIK665, S64315, AMG397 and AZD5991;

Aromatase inhibitors: exemestane (sold under the trade name Aromasin® from

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Pfizer), letrozole (sold under the trade name Femara® from Novartis), anastrozole (sold under the trade name Arimidex®);

Topoisomerase I inhibitors: irinotecan (marketed under the trade name Camptosar® from Pfizer), topotecan hydrochloride (marketed under the trade name Hycamtin® from GlaxoSmithKline);

Topoisomerase II inhibitors: etoposide (also known as VP-16 and etoposide phosphate, sold under the trade names Toposar®, VePesid® and Etopophos®), teniposide (also known as VM-26, sold under the trade name Vumon®);

mTOR inhibitors: temsirolimus (sold under the trade name Torisel® from Pfizer), ridaforolimus (formally known as deferolimus, (1R,2R,4S)-4-[(2R)2[(1R,9S,12S,15R,16E,18R ,19R,21R,23S,24E,26E,28Z,30S,32S,35R)-1,18-dihydroxy-19,30-dimethoxy15,17,21,23,29,35-hexamethyl-2,3,10, 14,20-pentaoxo-11,36-dioxa-4-azatricyclo[30.3,1,0 4' 9]hexatriaconta-16,24,26,28-tetraen-12-yl]propyl]-2-methoxycyclohexyl dimethylphosphinate, also known as AP23573 and MK8669 and described in PCT Publication No. WO 03/064383), everolimus (marketed under the trade name Afinitor® from Novartis);

A proteasome inhibitor such as carfilzomib, MLN9708, delanzomib, or bortezomib;

BET inhibitors such as INCB054329, OTX015, CPI-0610; LSD1 inhibitors such as GSK2979552, INCB059872; HIF-2a inhibitors such as PT2977 and PT2385;

Osteoclast bone resorption inhibitors: 1-hydroxy-2-imidazol-1ylphosphonoethyl)phosphonic acid monohydrate (sold under the trade name Zometa® from Novartis); CD33 antibody-drug conjugates: gemtuzumab ozogamicin (marketed under the trade name Mylotarg® from Pfizer/Wyeth);

CD22 antibody-drug conjugates: inotuzumab ozogamicin (also called CMC-544 and WAY-207294, available from Hangzhou Sage Chemical Co., Ltd.);

CD20 antibody-drug conjugates: ibritumomab tiutexan (marketed under the trade name Zevalin®);

Somatostatin analogues: octreotide (also known as octreotide acetate, sold under the trade names Sandostatin® and Sandostatin LAR®);

Synthetic interleukin-11 (IL-11): oprelvekin (marketed under the trade name Neumega® from Pfizer/Wyeth);

Synthetic erythropoietin: darbepoetin alfa (sold under the trade name Aranesp® from Amgen);

Receptor activator of nuclear factor B (RANK) inhibitors: denosumab (sold under the trade name Prolia® from Amgen);

Thrombopoietin mimetic peptibodies: romiplostim (sold under the trade name Nplate® from Amgen);

Cell growth promoters: palifermin (sold under the trade name Kepivance® from Amgen);

Antibodies against insulin-like growth factor-1 receptor (IGF-1R): figitumumab (also known as CP-751,871, available from ACC Corp), robatumumab (CAS No. 934235-44-6);

Anti-CSI antibodies: elotuzumab (HuLuc63, CAS No. 915296-00-3);

CD52 antibodies: alemtuzumab (sold under the trade name Campath®);

Histone deacetylase inhibitors (HDI): voninostat (sold under the trade name Zolinza® from Merck);

Alkylating agents: temozolomide (sold under the trade names Temodar® and Temodal® from Schering-Plough/Merck), dactinomycin (also known as actinomycin-D and sold under the trade name Cosmegen®), melphalan (also known as L-PAM, L- sarcolysine and phenylalanine mustard, sold under the trade name Alkeran®), altretamine (also known as hexamethylmelamine (HMM), sold under the trade name Hexalen®), carmustine (marketed under the trade name BiCNU®), bendamustine (marketed under the trade name Treanda®) , busulfan (sold under the trade names Busulfex® and Myleran®), carboplatin (sold under the trade name Paraplatin®), lomustine (also known as CCNU, sold under the trade name CeeNU®), cisplatin (also known as CDDP, sold under the trade names Platinol® and Platinol®-AQ), chlorambucil (sold under the trade name Leukeran®), cyclophosphamide (sold under the trade names Cytoxan® and Neosar®), dacarbazine (also known as DTIC, DIC and imidazole carboxamide, sold under the trade name DTIC- Dome®), altretamine (also known as hexamethylmelamine (HMM) sold under the trade name Hexalen®), ifosfamide (sold under the trade name Ifex®), procarbazine (sold under the trade name

- 31 044940

Matulane®), mechlorethamine (also known as nitrogen mustard, mustine and mechlorethamine hydrochloride, sold under the trade name Mustargen®), streptozocin (sold under the trade name Zanosar®), thiotepa (also known as thiophosphamide, TESPA and TSPA, sold under the trade name Thiopex®; biological response modifiers: Bacillus Calmette-Guérin (sold under the trade names theraCys® and TICE® BCG), denileukin diftitox (sold under the trade name Ontak®);

Antitumor antibiotics: doxorubicin (sold under the trade names Adriamycin® and Rubex®), bleomycin (sold under the trade name lenokcane®), daunorubicin (also known as daunorubicin hydrochloride, daunomycin and rubidomycin hydrochloride, sold under the trade name Cerubidine®), liposomal daunorubicin ( daunorubicin citrate liposomes, sold under the trade name DaunoKCome®), mitoxantrone (also known as DHAD, sold under the trade name Novantrone®), epirubicin (marketed under the trade name Ellence™), idarubicin (sold under the trade names Idamycin®, Idamycin PFS®) , mitomycin C (sold under the trade name Mutamycin®);

Agents that affect microtubules: estramustine (sold under the trade name Emcyl®);

Cathepsin K inhibitors: odanacatib (also known as MK-0822, N-(1-cyanocyclopropyl)-4fluoro-N2-{(1S)-2,2,2-trifluoro-1-[4'-(methylsulfonyl)biphenyl-4 -yl]ethyl}-L-leucinamide available from Lanzhou Chon Chemicals, ACC Corp. and ChemieTek and described in PCT Publication No. WO 03/075836); epothilone B analogues: ixabepilone (sold under the trade name Lxempra® from BristolMyers Squibb);

Heat shock protein (HSP) inhibitors: tanespimycin (17-allylamino-17demethoxygeldanamycin, also known as KOS-953 and 17-AAG, available from SIGMA and described in US Patent No. 4,261,989), NVP-HSP990, AUY922, AT13387, STA-9090 , Debio 0932, KW-2478, XL888, CNF2024, TAS-116.

TpoR agonists: eltrombopag (sold under the trade names Promacta® and Revolade® from GlaxoSmithKline);

Antimitotic agents: docetaxel (sold under the trade name Taxotere® from Sanofi-Aventis); adrenal steroid inhibitors: aminoglutethimide (sold under the trade name Cytadren®);

Antiandrogens: nilutamide (sold under the trade names Nilandron® and Anandron®), bicalutamide (sold under the trade name Casodex®), flutamide (sold under the trade name Fulexin™);

Androgens: fluoxymesterone (sold under the trade name Halotestin®);

Proteasome inhibitors: bortezomib (sold under the trade name Velcade®);

CDK inhibitors (CDK1, CDK2, CDK3, CDK5, CDK7, CDK8 or CDK9), including, but not limited to, alvocidib (a pan-CDK inhibitor, also known as flovopirdol or HMR-1275, 2-(2chlorophenyl)-5,7 -dihydroxy-8-[(3S,4R)-3-hydroxy-1-methyl-4-piperidinyl]-4-chromenone and described in US patent No. 5,621,002);

CDK4/6 inhibitors pabociclib, ribociclib, abemaciclib and trilaciclib; CDK9 inhibitors AZD 4573, P276-00, AT7519M, TP-1287;

Gonadotropin-releasing hormone receptor (GnRH) agonists: leuprolide or leuprolide acetate (marketed under the trade names Viadure® from Bayer AG, Eligard® from Sanofi-Aventis, and Lupron® from Abbott Lab);

Taxane antineoplastic agents: cabazitaxel (1-hydroxy-7,10-dimethoxy-9-oxo-5,20epoxytax-11-en-2a,4,13a-triyl-4-acetate-2-benzoate-13-[(2R, 3 S)-3 - {[(tert-butoxy)carbonyl]amino}-2hydroxy-3-phenylpropanoate), larotaxel ((2α,3ξ4α,5β,7α,10β,13α)-4,10-bis(acetyloxy)- 13-({(2R,3S)-3[(tert-butoxycarbonyl)amino]-2-hydroxy-3-phenylpropanoyl}oxy)-1-hydroxy-9-oxo-5,20-epoxy7,19-cyclotax-11 -en-2-yl benzoate);

5HTla receptor agonists: xaliprodene (also known as SR57746, 1-[2-(2-naphthyl)ethyl]-4-[3(trifluoromethyl)phenyl]-1,2,3,6-tetrahydropyridine and described in US Patent No. 5,266,573 ); LDC vaccines: Cervarix®, marketed by GlaxoSmithKline, Gardasil®, marketed by Merck; Iron chelating agents: deferazinox (sold under the trade name Exjade® OTNovartis);

Antimetabolites: claribine (2-chlorodeoxyadenosine, sold under the trade name leustatin®), 5-fluorouracil (sold under the trade name Adrucil®), 6-thioguanine (sold under the trade name Purinethol®), pemetrexed (sold under the trade name Alimta®), cytarabine (also known as arabinosylcytosine (Ara-C), sold under the trade name Cytosar-U®), liposomal cytarabine (also known as liposomal Ara- 32 044940

C, sold under the trade name DepoCyt™), decitabine (sold under the trade name Dacogen®), hydroxyurea (sold under the trade names Hydrea®, Droxia™ and Mylocel™), fludarabine (sold under the trade name Fludara®), floxuridine (sold under the trade name Fludara®), floxuridine (sold under the trade name Fludara®), trade name FUDR®), cladribine (also known as 2-chlorodeoxyadenosine (2-CdA) sold under the trade name Leustatin™), methotrexate (also known as amethopterin, methotrexane sodium (MTX) sold under the trade names Rheumatrex® and Trexall™), pentostatin (sold under the trade name Nipent®);

Bisphosphonates: pamidronate (sold under the trade name Aredia®), zoledronic acid (sold under the trade name Zometa®); demethylating agents: 5-azacytidine (sold under the trade name Vidaza®), decitabine (sold under the trade name Dacogen®);

Plant alkaloids: protein-bound paclitaxel (sold under the trade name Abraxane®), vinblastine (also known as vinblastine sulfate, vincaleucoblastine and VLB, sold under the trade names Alkaban-AQ® and Velban®), vincristine (also known as vincristine sulfate, LCR and VCR, sold under the trade names Oncovin® and Vincasar Pfs®), vinorelbine (sold under the trade name Navelbine®), paclitaxel (sold under the trade names Taxol and Onxal™);

Retinoids: ali tretinoin (sold under the trade name Panretin®), tretinoin (polytransretinoic acid, also known as ATRA, sold under the trade name Vesanoid®), isotretinoin (13-cis-retinoic acid, sold under the trade names Accutane®, Amnesteem®, Claravis®, Clams®, Decutan®, Isotane®, Izotech®, Oratane®, Isotret® and Sotret®), bexarotene (sold under the trade name Targretin®);

Glucocorticosteroids: hydrocortisone (also known as cortisone, hydrocortisone sodium succinate, hydrocortisone sodium phosphate, and sold under the trade names Ala-Cort®, Hydrocortisone Phosphate, Solu-Cortef®, Hydrocort Acetate® and Lanacort®), dexamethasone ((8S,9R, 10S,11S,13S,14S,16R,17R)fluoro-11,17-dihydroxy-17-(2-hydroxyacetyl)-10,13,16-trimethyl-6,7,8,9,10,11,12, 13,14,15,16,17-dodecahydro3H-cyclopenta[a]phenanthrene-3-one), prednisolone (sold under the trade names DeltaCortel®, Orapred®, Pediapred® and Prelone®), prednisone (sold under the trade names Deltasone® , Liquid Red®, Meticorten® and Orasone®), methylprednisolone (also known as 6methylprednisolone, methylprednisolone acetate, methylprednisolone sodium succinate, sold under the trade names Duralone®, Medralone®, Medrol®, M-Prednisol® and Solu-Medrol®);

Cytoxins: interleukin-2 (also known as aldesleukin and IL-2, sold under the trade name Proleukin®), interleukin-11 (also known as oprevelkin, sold under the trade name Neumega®), interferon alpha (also known as IFN-alpha , sold under the trade names Intron® A and Roferon-A®); [00209] estrogen receptor inhibitors: fulvestrant (sold under the trade name Faslodex®);

Antiestrogens: tamoxifen (sold under the trade name Novaldex®); toremifene (sold under the trade name Fareston®);

Selective estrogen receptor modulators (SERMs): raloxifene (sold under the trade name Evista®);

Luteinizing hormone-releasing hormone (LHRH) agonists: goserelin (sold under the trade name Zoladex®); progesterones: megestrol (also known as megestrol acetate, sold under the trade name Megace®);

Miscellaneous cytotoxic agents: arsenic trioxide (sold under the trade name Trisenox®), asparaginase (also known as L-asparaginase, Erwinia L-asparaginase, sold under the trade names Elspar® and Kidrolase®);

One or more additional immune checkpoint inhibitors may be used in combination with a compound as described herein to treat SHP2-associated diseases, disorders or conditions. Typical immune checkpoint inhibitors include inhibitors (smack molecules or biologics) of immune checkpoint molecules such as CD27, CD28, CD40, CD122, CD96, CD73, CD39, CD47, OX40, GITR, CSF1R, JAK, PI3K delta, PI3K gamma , TAM kinase, arginase, CD137 (also known as 4-1BB), ICOS, A2AR, A2BR, HIF-2a,

B7-H3, B7-H4, BTLA, CTLA-4, LAG3, TIM3, VISTA, CD96, TIGIT, PD-1, PD-L1 and PD-L2. In some embodiments, the immune checkpoint molecule is a stimulatory immune checkpoint molecule selected from CD27, CD28, CD40, ICOS, OX40, GITR, CD137, and STING. In some embodiments, the immune checkpoint molecule is an inhibitory immune checkpoint molecule selected from B7-H3, B7-H4, BTLA, CTLA-4, IDO, TDO, arginase, KIR, LAG3, PD-1, TIM3, CD96, TIGIT and VISTA. In some embodiments, the compounds provided herein may be used in combination with one or more agents selected from KIR inhibitors, TIGIT inhibitors, LAIR1 inhibitors, CD160 inhibitors, 2B4 inhibitors and TGFR beta inhibitors.

In some embodiments, the immune checkpoint molecule inhibitor is a PD-1 inhibitor, such as an anti-PD-1 monoclonal antibody. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab, pembrolizumab (also known as MK-3475), pidilizumab, SHR-1210, PDR001, or AMP-224. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab or pembrolizumab or PDR001. In some embodiments, the anti-PD1 antibody is pembrolizumab.

In some embodiments, the immune checkpoint molecule inhibitor is a PD-L1 inhibitor, such as an anti-PD-L1 monoclonal antibody. In some embodiments, the αntu-PD-L1 monoclonal antibody is BMS-935559, MEDI4736, MPDL3280A (also known as RG7446), or MSB0010718C. In some embodiments, the anti-PD-L1 monoclonal antibody is MPDL3280A (atezolizumab) or MEDI4736 (durvalumab).

In some embodiments, the immune checkpoint molecule inhibitor is a CTLA-4 inhibitor, such as an anti-CTLA-4 antibody. In some embodiments, the anti-CTLA-4 antibody is ipilimumab or tremelimumab. In some embodiments, the immune checkpoint molecule inhibitor is a LAG3 inhibitor, such as an anti-LAG3 antibody. In some embodiments, the anti-LAG3 antibody is BMS-986016 or LAG525. In some embodiments, the immune checkpoint molecule inhibitor is a GITR inhibitor, such as an anti-GITR antibody. In some embodiments, the anti-GITR antibody is TRX518 or MK-4166, INCAGN01876 or MK-1248. In some embodiments, the immune checkpoint molecule inhibitor is an OX40 inhibitor, such as an anti-OX40 antibody or an OX40L fusion protein. In some embodiments, the anti-OX40 antibody is MEDI0562 or INCAGN01949, GSK2831781, GSK-3174998, MOXR-0916, PF04518600, or LAG525. In some embodiments, the OX40L fusion protein is MEDI6383.

The compounds of the invention can also be used to enhance or enhance the immune response, including enhancing the immune response to an antigen; to improve immunization, including increasing vaccine effectiveness; and to increase inflammation. In some embodiments, the compounds of the invention may be used to improve the immune response to vaccines, including, but not limited to, Listeria vaccines, oncolytic virus vaccines, and cancer vaccines such as GVAX® (granulocyte-macrophage colony-stimulating factor gene-transfected (GM-CF) ) antitumor cell vaccine). Cancer vaccines include dendritic cells, synthetic peptides, DNA vaccines, and recombinant viruses. Other immunomodulatory agents also include those that block immune cell migration, such as chemokine receptor antagonists, including CCR2 and CCR4; STING agonists and Toll receptor agonists.

Other anticancer agents also include those that enhance the immune system, such as adjuvants or T cell adoptive transfer. The compounds of the present invention may be effective in combination with CAR (chimeric antigen receptor) T cell treatment as an enhancer of T cell activation.

The compound of the present invention can also be used in combination with the following adjuvant therapies:

Antiemetic drugs: NK-1 receptor antagonists: casopitant (marketed under the trade names Rezonic® and Zunrisa® from GlaxoSmithKline); And

Cytoprotectors: amifostine (sold under the trade name Ethyol®), leucovorin (also known as leucovorin calcium, citrovorum factor and folinic acid).

Examples

The following methods for preparing intermediates (standards) of compounds of formula (IIB) are provided to enable those skilled in the art to more clearly understand and apply the present disclosure in practice. They should not be construed as limiting the scope of the description, but merely as illustrative and typical.

Standard 1.

Synthesis of (5S)-5,6-dihydrospiro[piperidine-4,4-pyrrolo[1,2-b]pyrazole]-5-amine dihydrochloride.

Step 1: 1-[1-[(tert-butoxy)(hydroxy)methyl]-4-[hydroxy(methoxy)methyl]piperidin-4-yl]ethan-1-ol.

- 34 044940

To a solution of [1-[(tert-butoxy)(hydroxy)methyl]piperidin-4-yl](methoxy)methanol (20 g, 80.86 mmol, 1.0 equiv) in THF (200 ml) at -78° Add LDA (48.52 mL, 97.03 mmol, 1.2 equiv) dropwise under nitrogen. After stirring for 1.5 hours at -78°C to 60°C, acetaldehyde (5.34 g, 121.29 mmol, 1.5 equiv) was added dropwise over 5 minutes at -78°C. and the resulting mixture is stirred for an additional 2 hours at -78°C -40°C. The reaction mixture is then poured into sat. aq. NH ₄ Cl solution and extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography (EtOAc/PE, 0-50%) to give 12 g (50.9% yield) of the title compound as a light yellow oil.

Stage 2: dicarboxylate.

1-tert-butyl 4-methyl 4-[1-(trifluoromethanesulfonyloxy)ethyl]piperidine-1,4-

To a solution of 1-tert-butyl 4-methyl 4-(1-hydroxyethyl)piperidine-1,4-dicarboxylate (9.5 g, 33.06 mmol, 1 equiv) in DCM (100 ml) add pyridine (10.4 g, 131.48 mmol, 4.0 equiv) and (trifluoromethane)sulfonyl trifluoromethanesulfonate (18.7 g, 66.27 mmol, 2.0 equiv) dropwise at 0°C under nitrogen atmosphere. After stirring for 2 hours at 0°C, the reaction mixture is quenched with water at 0°C. The resulting mixture is extracted with CH2Cl2. The organic layer was washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure to obtain 20 g (crude) of the title compound. This crude product is used directly in the next step without further purification.

Step 3: 1-tert-butyl 4-methyl 4-ethenylpiperidine-1,4-dicarboxylate.

To a stirred solution of 1-tert-butyl 4-methyl 4-[1-(trifluoromethanesulfonyloxy)ethyl]piperidine-1,4-dicarboxylate (20 g, crude) in DCM (300 ml) add DBU (28.50 ml, 187, 18 mmol) at rt in a nitrogen atmosphere. After stirring for 4 hours at rt, the reaction mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography (EtOAc/PE, 6%) to give the title compound (4.5 g, 55.2% yield in two steps ) as a light yellow oil.

Step 4: 1-[(tert-butoxy)carbonyl]-4-ethenylpiperidine-4-carboxylic acid.

To a solution of 1-tert-butyl 4-methyl 4-ethenylpiperidine-1,4-dicarboxylate (4.8 g, 17.82 mmol, 1.0 equiv) in MeOH (40 ml) add water (10 ml) and LiOH ( 2.35 g, 98.01 mmol, 5.5 equiv) at room temperature. After stirring for 16 hours at rt, the mixture is acidified to pH=5 with 0.5 M HCl aq. solution. The reaction mixture is then extracted with CH ₂ Cl ₂ . The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and concentrated to give the title compound (4.5 g, 98.9% yield) as a light yellow oil, which was used in the next step without further purification.

Step 5: tert-butyl 4-ethenyl-4-[methoxy(methyl)carbamoyl]piperidine-1-carboxylate.

To a solution of 1-[(tert-butoxy)carbonyl]-4-ethenylpiperidine-4-carboxylic acid (4.5 g, 17.62 mmol,

- 35 044940

1.0 equiv) and methoxy(methyl)amine (1.61 g, 26.43 mmol, 1.5 equiv) in DCM (70 ml) add HATU (13.40 g, 35.25 mmol, 2.0 equiv ) and Et ₃ N (5.35 g, 52.86 mmol, 3.0 equiv) at rt under nitrogen. After stirring for 12 hours at rt, the reaction mixture was quenched with water at room temperature and extracted with DCM. The organic layer was washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography (EtOAc/PE, 5%) to give the title compound (5 g, 95% yield) as a white solid.

Step 6: tert-butyl 4-acetyl-4-ethenylpiperidine-1-carboxylate.

To a solution of tert-butyl 4-ethenyl-4-[methoxy(methyl)carbamoyl]piperidine-1-carboxylate (5 g, 16.75 mmol, 1.0 equiv) in THF (60 ml) add 2.5 M CH ₃ MgBr (16.76 mL, 41.89 mmol, 2.50 equiv) dropwise at 0°C under nitrogen. After stirring for 12 hours at rt, the reaction is quenched with sat. aq. NH ₄ Cl solution at 0°C and the mixture is extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography (EtOAc/PE, 1/3) to give the title compound (3.5 g, 82.4% yield) as a light yellow oil.

Step 7: tert-butyl 4-[(2Z)-3-(dimethylamino)prop-2-enoyl]-4-ethenylpiperidine-1-carboxylate.

A solution of tert-butyl 4-acetyl-4-ethenylpiperidine-1-carboxylate (2.2 g, 8.68 mmol, 1.0 equiv) in [(tert-butoxy)(dimethylamino)methyl]dimethylamine (2 ml, 9. 69 mmol, 1.1 equiv.) is stirred for 4 hours at 100°C under nitrogen atmosphere. The reaction mixture is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE/EtOAc (1:1) to give the title compound (1.6 g, 59.7% yield) as a light yellow oil.

Step 8: tert-butyl 4-ethenyl-4-(1H-pyrazol-5-yl)piperidine-1-carboxylate.

a solution of tert-butyl 4-[(2Z)-3-(dimethylamino)prop-2-enoyl]-4K stirred ethenylpiperidine-1-carboxylate (2.2 g, 7.13 mmol, 1.0 equiv.) in EtOH ( 50 ml) add hydrazine monohydrate (0.54 g, 10.78 mmol, 1.5 equiv) at 25°C under nitrogen atmosphere. After stirring for 16 hours at 25°C, the reaction is quenched with water and the resulting mixture is extracted with DCM. The organic layer was washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (30-60%) to give the title compound (1.6 g, 80.8% yield) as a white solid.

Step 9: tert-butyl 4-(oxiran-2-yl)-4-(1H-pyrazol-5-yl)piperidine-1-carboxylate.

To a stirred mixture of tert-butyl 4-ethenyl-4-(1H-pyrazol-5-yl)piperidin-1-carboxylate (2 g, 7.2 mmol, 1.0 equiv) and methyltrioxorhenium(VI) (179.72 mg , 0.72 mmol, 0.1 equiv) to DCM (30 ml) add pyridine (228.14 mg, 2.88 mmol, 0.4 equiv) and H ₂ O ₂ (30%) (1.23 g, 10.85 mmol, 1.5 equiv) at ct. After stirring for 16 hours at rt, the reaction is quenched with water and extracted with DCM. The organic layer was washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (30-60%) to give the title compound (0.8 g, 37.82%) as a white solid.

Step 10: tert-butyl 5-hydroxy-5,6-dihydrospiro[piperidine-4,4-pyrrolo[1,2-b]pyrazole] - 36 044940

-carboxylate.

To a stirred solution of tert-butyl 4-(oxiran-2-yl)-4-(1H-pyrazol-5-yl)piperidine-1carboxylate (0.7 g, 2.38 mmol, 1.0 equiv) in THF (10 ml) add LiBr (0.62 g, 7.15 mmol, 3.0 equiv) and CH3COOH (0.43 g, 7.15 mmol, 3.0 equiv) at room temperature. After stirring for 16 hours at rt, the reaction mixture was stirred at 45°C for 8 hours. After cooling to rt, the reaction mixture was quenched with sat. aq. NaHCO ₃ solution and the mixture is extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (30-100%) to give the title compound (0.4 g, 57.1% yield) as a white solid.

Step 11: tert-butyl 5-oxo-5,6-dihydrospiro[piperidine-4,4-pyrrolo[1,2-b]pyrazole]-1carboxylate.

To a solution of tert-butyl 5-hydroxy-5,6-dihydrospiro[piperidine-4,4-pyrrolo[1,2-b]pyrazole]-1carboxylate (0.4 g, 1.36 mmol, 1.0 equiv) in DCM (8 ml) add Dess-Martin reagent (0.87 g, 2.04 mmol, 1.5 equiv) at rt. After stirring for 4 hours at room temperature, the reaction mixture is quenched with sat. aq. NaHCO ₃ solution at room temperature and extracted with DCM. The organic layer was washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by silica gel column chromatography (EtOAc/PE, 20-60%) to give the title compound (0.36 g, 90.62%) as a white solid.

Step 12: tert-butyl (S)-5-[(R)-2-methylpropane-2-sulfinyl)amino]-5,6-dihydrospiro[piperidine4,4-pyrrolo[1,2-b]pyrazole] -1 -carboxylate.

To a solution of tert-butyl 5-oxo-5,6-dihydrospiro[piperidine-4,4-pyrrolo[1,2-b]pyrazole]-1carboxylate (0.36 g, 1.23 mmol, 1.0 equiv.) in THF (6 ml) add (R)-2-methylpropane-2-sulfinamide (0.30 g, 2.47 mmol, 2.0 equiv.) and Ti(OEt) ₄ (1.13 g, 4.94 mmol, 4.0 equiv.) at rt in a nitrogen atmosphere. After stirring for 4 hours at 75°C, the reaction mixture is cooled to -20°C. To the resulting mixture add MeOH (1 ml) and LiBH ₄ (40.38 mg, 1.85 mmol, 1.50 equiv) at -20°C. After stirring for another 6 hours at 0°C, the reaction mixture is quenched with sat. aq. NH4C1 solution at 0°C. The mixture is then filtered and the filter cake is washed with EtOAc. The filtrate is concentrated under reduced pressure and the residue is dissolved in MeOH (3 ml). LiBH ₄ (80.75 mg, 3.71 mmol, 3.0 equiv) was added to the resulting mixture at room temperature. The resulting mixture was stirred for an additional 8 hours at 45°C. The reaction mixture is quenched with sat. aq. NH4C1 solution at 0°C. The resulting mixture is extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (30%-70%) to give the title compound (200 mg, 40.8% yield) as a white solid.

Step 13: (5S)-5,6-dihydrospiro[piperidine-4,4-pyrrolo[1,2-b]pyrazole]-5-amine dihydrochloride.

To a stirred solution of tert-butyl (S)-5-[(R)-2-methylpropane-2-sulfinyl)amino]-5,6dihydrospiro[piperidine-4,4-pyrrolo[1,2-b]pyrazole]-1 -carboxylate (100 mg, 0.252 mmol, 1.0 equiv) in 1,4-dioxane (0.5 ml), add a solution of HCl in dioxane (4 M, 0.50 ml) dropwise at room temperature. After stirring for 30 minutes at rt, the reaction mixture is concentrated under reduced pressure. Et2O (1 mL) was added to the residue and the precipitate was collected by filtration to obtain the title compound (60 mg, 89.7% yield) as a white solid.

Example 1

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((S)-6a,7, 8,9-tetrahydro6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol.

Step 1: tert-butyl (2S)-2-[[(2-fluoropyridin-3-yl)oxy]methyl]pyrrolidine-1-carboxylate.

To a stirred mixture of 2-fluoropyridin-3-ol (1 g, 8.84 mmol, 1.00 equiv.) and tert-butyl (2S)-2(hydroxylmethyl)pyrrolidin-1-carboxylate (2.14 g, 10. 61 mmol, 1.2 equiv.) in THF (15 ml) add PPh ₃ (3.48 g, 13.26 mmol, 1.50 equiv) and DEAD (2.31 g, 13.26 mmol, 1.5 equiv.) at room temperature in a nitrogen atmosphere. After stirring for 16 hours at room temperature, the reaction mixture is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE/EtOAc (15:1) to give the title compound (2.5 g, 95.4% yield) as a light yellow oil.

Step 2: 2-fluoro-3-[[(2S)-pyrrolidin-2-yl]methoxy]pyridine dihydrochloride.

To a stirred solution of tert-butyl (2S)-2-[[(2-fluoropyridin-3-yl)oxy]methyl]pyrrolidine-1-carboxylate (2500 mg, 8.43 mmol, 1.00 equiv) in DCM (15 ml) add hydrogen chloride (4M in dioxane) (15 ml) at room temperature. After stirring at rt for 4 hours, the reaction mixture was concentrated in vacuo to give the title compound (2.20 g, 96.8% yield) as a white solid.

Step 3: (S)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazine.

K2CO3 ( 5.64 g, 40.87 mmol, 5.00 equiv) at room temperature, and the resulting mixture was stirred for 12 hours at 65°C. The mixture is then filtered and the filtrate is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE/EtOAc (3:1) to give the title compound (1.34 g, 93% yield) as a colorless oil.

Step 4: (S)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazine.

To a stirred solution of (S)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazine (600 mg, 3.41 mmol, 1.00 equiv) in THF (15 ml) add dropwise a solution of n-butyllithium (2.5 M in hexane, 3.4 ml, 8.5 mmol, 2.50 equiv) at -78°C under N2. The reaction mixture is heated to 0°C and stirred for 1.5 hours. A solution of I2 (950.60 mg, 3.74 mmol, 1.10 equiv) in THF (2 ml) is added dropwise at -78 °C. The resulting mixture is warmed to room temperature and stirred for 2 hours at room temperature. The reaction mixture is quenched with sat. aq. NH ₄ Cl solution and diluted with EtOAc. The organic layer was washed with brine, dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE/EtOAc (3:1) to give the title compound (700 mg, 68% yield) as a yellow solid.

Step 5: 2-ethylhexyl yl)thio)propanoate.

3-(((8)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-0][1,4]oxazin-4-

To a stirred mixture of (8)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2<1][1,4]oxazine (150.00 mg , 0.496 mmol, 1.00 equiv), 2-ethylhexyl 3-sulfanylpropanoate (162.62 mg, 0.745 mmol, 1.50 equiv), Pd ₂ (dba) ₃ (22.73 mg, 0.025 mmol, 0.05 equiv ) and Xantphos (14.36 mg, 0.025 mmol, 0.05 equiv) in 1,4-dioxane (2 ml) add DIEA (192.51 mg, 1.489 mmol, 3.00 equiv) at room temperature under nitrogen atmosphere. After stirring for 1 hour at 90°C, the reaction mixture is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-30%) to give the title compound (160 mg, 82% yield) as a light yellow solid.

Step 6: (8)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-0][1,4]oxazine-4-thiolate potassium.

To a stirred solution of 2-ethylhexyl 3-(((8)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-0][1,4]oxazine-4 -yl)thio)propanoate (140.00 mg, 0.357 mmol, 1.0 equiv) in THF (2 ml) add 1.0 M t-ViOA (0.43 ml, 0.43 mmol, 1.20 equiv) at -10°C. After stirring for 0.5 h at 0°C, the reaction mixture is diluted with petroleum ether. The precipitated solid was collected by filtration and washed with ethyl acetate to give the title compound (75 mg, 85% yield) as a light yellow solid.

Step 7: Methyl 6-bromo-3-[(38.48)-4-[[(tert-butoxy)carbonyl]amino]-3-methyl-2-oxa-8azaspiro[4,5]decan-8-yl ]pyrazine-2-carboxylate.

Solution of methyl 3,6-dibromopyrazine-2-carboxylate (500 mg, 1.690 mmol, 1 equiv), (38.48)-3-methyl-2oxa-8-azaspiro[4,5]decane-4-amine dihydrochloride (493 .05 mg, 2.028 mmol, 1.2 equiv) and DIEA (1091.88 mg,

8.448 mmol, 5.0 equiv) in DMA (10 ml) was stirred for 2 hours at 55°C. Di-tert-butyl dicarbonate (552.33 mg, 2.531 mmol, 1.5 equiv) was then added and the resulting mixture was stirred for 2 hours at room temperature. The reaction mixture is diluted with water and extracted with EtOAc. The organic layer was washed with water and brine, dried over anhydrous No. ₂ 8O | and concentrate. The residue was purified by silica gel column chromatography, eluting with PE/EtOAc=1:1 to give the title compound (615 mg, 2 steps 75% yield) as a yellow oil.

Step 8: tert-butyl M-[(38.48)-8-[5-bromo-3-(hydroxymethyl)pyrazin-2-yl]-3-methyl-2-oxa-8azaspiro[4.5] decane- 4 -yl] carbamate.

To the stirred solution methyl 6-bromo-3-[(38.48)-4-[[(tert-butoxy)carbonyl]amino]-3-methyl2-oxa-8-azaspiro[4,5]decan-8-yl ]pyrazine-2-carboxylate (150 mg, 0.309 mmol, 1 equiv) in DCM (3.75 ml) add DIBAL-H (1.0 M in DCM, 1.24 ml, 1.240 mmol, 4.01 equiv) at drops at -78°C in a nitrogen atmosphere. The resulting mixture is stirred for 30 minutes at -78°C in a nitrogen atmosphere, heated to rt and then quenched by adding sat. aq. potassium sodium tartrate solution. The resulting mixture is extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous

-39044940

Na2SO ₄ and concentrate. The residue is purified by column chromatography on silica gel, eluted

EtOAc/PE (0-50%) to give the title compound (65 mg, 46%) as a yellow solid.

Step 9: tert-butyl ((3S,4S)-8-(3-(hydroxymethyl)-5-(((S)-6a,7,8,9-tetrahydro-6H-pyrido[3,2b]pyrrolo[ 1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4,5]decan-4-yl)carbamate.

To the stirred mixture tert-butyl N-[(3S,4S)-8-[5-bromo-3-(hydroxymethyl)pyrazin-2-yl]-3methyl-2-oxa-8-azaspiro[4,5]decane- 4-yl]carbamate (25.00 mg, 0.055 mmol, 1.00 equiv), (S)-6a,7,8,9tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d] Potassium [1,4]oxazine-4-thiolate (20.20 mg, 0.082 mmol, 1.50 equiv), Pd ₂ (dba) ₃ (15.02 mg, 0.0164 mmol, 0.30 equiv) and Xantphos (9.49 mg, 0.0164 mmol, 0.30 equiv) in 1,4dioxane (1 ml) add DIEA (21.19 mg, 0.164 mmol, 3.00 equiv) at room temperature under nitrogen. After stirring for 1.5 hours at 100°C, the reaction mixture is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (50-100%) to give the title compound (25 mg, 78% yield) as a light yellow solid.

Step 10: (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((S)-6a, 7,8,9tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d] [1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol formate.

To a stirred solution tert-butyl ((3S,4S)-8-(3-(hydroxymethyl)-5-(((S)-6a,7,8,9-tetrahydro-6Hpyrido[3,2-b]pyrrolo[ 1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4,5]decane-4yl)carbamate (20, 00 mg, 0.034 mmol, 1.0 equiv) in DCM (1.5 ml) add TFA (0.3 ml) at room temperature. After stirring for 5 hours at room temperature, the reaction mixture is concentrated under reduced pressure. The crude product was purified by pre-HPLC to give the title compound (7 mg, 39% yield) as a light yellow solid. MS (ER, m/z): [M+1]+ = 485.2.

Example 2.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((R)-6a,7, 8,9-tetrahydro6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol.

Compound 2 was synthesized by the method described in Example 1 using tert-butyl (2R)-2(hydroxymethyl)pyrrolidine-1-carboxylate instead of tert-butyl (2S)-2-(hydroxylmethyl)pyrrolidine-1-carboxylate in step 1. MS ( ER, m/z): [M+1] ⁺ = 485.3.

Example 3.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)-8- fluoro-6a,7,8,9tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol.

Step 1: tert-butyl (2S,4S)-4-fluoro-2-(hydroxymethyl)pyrrolidine-1-carboxylate.

- 40 044940

To a stirred solution of (2S,4S)-1-(tert-butoxycarbonyl)-4-fluoropyrrolidine-2-carboxylic acid (2.33 g, 9.99 mmol, 1.0 equiv) in THF (25 ml) add BH ₃ -Me ₂ S (2.8 ml, 29.5 mmol, 3.0 equiv) was dropped at 0-5° C. under nitrogen and the resulting mixture was stirred overnight at room temperature. The mixture is cooled to 0°C and quenched with MeOH. The resulting mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography, eluting with EtOAc/PE (1/1) to give the title compound (2.0 g, 91.3%).

Step 2: tert-butyl (2S,4S)-4-fluoro-2-[[(2-fluoro-4-iodopyridin-3-yl)oxy]methyl]pyrrolidine-1-carboxylate.

To a stirred solution of 2-fluoro-4-iodopyridin-3-ol (436 mg, 1.82 mmol, 1.0 equiv), tert-butyl (2S,4S)-4-fluoro-2-(hydroxymethyl)pyrrolidine-1-carboxylate (400 mg, 1.82 mmol, 1.0 equiv) and PPh ₃ (717 mg, 2.73 mmol, 1.5 equiv) in THF (4.00 ml) add DEAD (476 mg, 2.73 mmol, 1.5 equiv) dropwise at 0°C in a nitrogen atmosphere. The resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere. The reaction mixture is quenched with water and the resulting mixture is extracted with EtOAc. The combined organic layers are washed with water and brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-50%), to give the title compound (610 mg, 75.5%).

Step 3: 2-fluoro-3-(((2S,4S)-4-fluoropyrrolidin-2-yl)methoxy)-4-iodopyridine hydrochloride.

To a stirred solution of tert-butyl (2S,4S)-4-fluoro-2-[[(2-fluoro-4-iodopyridin-3yl)oxy]methyl]pyrrolidine-1-carboxylate (560 mg, 1.272 mmol, 1.0 equiv) in 1,4-dioxane (5 ml) add a solution of 3.3 M HCl in 1,4-dioxane (5 ml, 16.5 mmol, 13 equiv) dropwise at 0°C. After stirring at room temperature for 1 hour, the reaction mixture was concentrated under reduced pressure and the residue triturated with Et ₂ O. The solid was collected by filtration and dried in vacuo to give the product as the HCl salt (375 mg, 78.3%).

Step 4: (6aS,8S)-8-fluoro-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4] oxazine

A mixture of 2-fluoro-3-(((2S,4S)-4-fluoropyrrolidin-2-yl)methoxy)-4-iodopyridine hydrochloride (370 mg, 0.983 mmol, 1.0 equiv) and K2CO3 (407 mg, 2.0 equiv) 95 mmol, 3.0 equiv) in EtOH (4 ml) was stirred for 2 hours at 60°C. The resulting mixture is diluted with water and extracted with EtOAc. The combined organic layers are washed with water and brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-30%) to give the title compound (245 mg, 77.9%).

Step 5: 2-ethylhexyl 3-([5-[(3S,4S)-4-[(tert-butoxycarbonyl)amino]-3-methyl-2-oxa-8azaspiro[4,5]decan-8-yl] -6-(hydroxymethyl)pyrazin-2-yl]sulfanyl)propanoate.

A solution of tert-butyl N-[(3S,4S)-8-[5-bromo-3-(hydroxymethyl)pyrazin-2-yl]-3-methyl-2-oxa-8azaspiro[4,5]decane-4- yl]carbamate (580.0 mg, 1.27 mmol, 1.0 equiv), 2-ethylhexyl 3sulfanylpropanoate (332.29 mg, 1.52 mmol, 1.2 equiv), Pd ₂ (dba) ₃ (116, 1 mg, 0.127 mmol, 0.10 equiv), Xantphos (73.38 mg, 0.127 mmol, 0.10 equiv) and DIEA (491.69 mg, 3.804 mmol, 3.00 equiv) in 1,4-dioxane

- 41 044940 (12.0 ml) is stirred for 1 hour at 100°C in a nitrogen atmosphere. The resulting mixture was concentrated in vacuo and the residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-50%) to give the title compound (600 mg, 79.54%).

Step 6: 5-((38.48)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4,5]-decan-8-yl)6-(hydroxymethyl )sodium pyrazine-2-thiolate.

To a stirred solution of 2-ethylhexyl 3-([5-[(38.48)-4-[(tert-butoxycarbonyl)amino]-3 methyl-2-oxa-8-azaspiro[4,5]decan-8-yl ]-6-(hydroxymethyl)pyrazin-2-yl]sulfanyl)propanoate (680.0 mg, 1.14 mmol, 1.0 equiv) in CH ₃ OH (6.80 ml) add CH ₃ ONa (247.04 mg, 1.372 mmol, 1.20 equiv, 30% in MeOH) dropwise at 5°C. After stirring at rt for 16 hours, the reaction mixture was concentrated in vacuo and the residue triturated with Et ₂ 0c to give the title compound (390 mg, crude), which was used in the next step without further purification.

Step 7: tert-butyl ((38.48)-8-(5-(((6a8.88)-8-fluoro-6a,7,8,9-tetrahydro-6H-pyrido[3,2b]pyrrolo[ 1,2-b][1,4]oxazin-4-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4,5]decane- 4-yl)carbamate.

Solution of (6a8,88)-8-fluoro-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-b][1,4]oxazine ( 30.00 mg, 0.094 mmol, 1.0 equiv), 5-((38.48)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8azaspiro[4,5]decane- Sodium 8-yl)-6-(hydroxymethyl)pyrazine-2-thiolate (48.64 mg, 0.112 mmol, 1.2 equiv), Pd ₂ (dba) ₃ (25.75 mg, 0.028 mmol, 0.30 equiv ), Xantphos (16.27 mg, 0.028 mmol, 0.30 equiv) and DIEA (36.34 mg, 0.281 mmol, 3.00 equiv) in dioxane (0.90 ml) were stirred for 1 hour at 80°C in a nitrogen atmosphere. After cooling to rt, the reaction mixture is diluted with water and extracted with EtOAc. The organic layer is washed with water and brine, dried over Na ₂ SO4 and concentrated. The residue was purified by silica gel column chromatography, eluting with MeOH/CH ₂ Cl ₂ (0-10%) to give the title compound (32 mg, 56.4%).

Step 8: (3-((8)-4-amino-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6a8,88)-8-fluoro-6a, 7,8,9-tetrahydro-6Hpyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol.

To a solution of tert-butyl ((38.48)-8-(5-(((6a8.88)-8-fluoro-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo [1,2b][1,4]oxazin-4-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4,5]decan-4yl )carbamate (26 mg, 0.043 mmol, 1.00 equiv) in DCM (1.2 ml) add TFA (0.4 ml, 5.23 mmol, 122 equiv) dropwise at 5°C. After stirring at rt for 1 hour, the reaction mixture was concentrated in vacuo and the residue was purified by pre-HPLC to give the title compound (3.1 mg, 14.30%). MS (ER, m/z): [M+1] ⁺ = 503.2.

Example 4.

Synthesis of (3-((38.48)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(((6a8.8K)-8- fluoro-6a,7,8,9tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-b][1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol.

-42 044940

The title compound was synthesized by the method described in Example 3 using (2S,4R)-1-(tert-butoxycarbonyl)-4-fluoropyrrolidine-2-carboxylic acid in step 1. MS (ES, m/z): [M +1]+ = 503.3.

Example 5.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)-8- methoxy-6a,7,8,9tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol.

Step 1: 1-tert-butyl dicarboxylate.

2-methyl (2S,4S)-4-[(tert-butyldimethylsilyl)oxy]pyrrolidine-1,2-

solution of 1-tert-butyl 2-methyl

To stirred (2S,4S)-4-hydroxypyrrolidine-1,2 dicarboxylate (6.0 g, 24.46 mmol, 1.0 equiv) and imidazole (2.5 g, 36.72 mmol, 1.50 equiv) in DCM (60 ml) add TBS-Cl (5.53 g, 36.69 mmol, 1.50 equiv) in portions at 0-5°C. After stirring overnight at room temperature, the reaction mixture was quenched with MeOH and water, and the resulting mixture was extracted with EtOAc. The combined organic layers are washed with water and brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-10%) to give the title compound (9.0 g, 102.3%).

Step 2: tert-butyl (2S,4S)-4-[(tert-butyldimethylsilyl)oxy]-2-(hydroxymethyl)pyrrolidine-1carboxylate.

To a stirred mixture of 1-tert-butyl 2-methyl (2S,4S)-4-[(tert-butyldimethylsilyl)oxy]pyrrolidine-1,2-dicarboxylate (3.0 g, 8.344 mmol, 1.0 equiv) in THF (30 ml) add LiAlH ₄ (475 mg,

12.516 mmol, 1.50 equiv) portions at 0° C. under a nitrogen atmosphere, and the resulting mixture was stirred for 2 hours at room temperature under a nitrogen atmosphere. After cooling to 0°C, the reaction mixture is quenched with sat. aq. Na ₂ SO ₄ solution. The resulting mixture is filtered and the filtrate is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-100%), to give the title compound (1.6 g, 57.8%).

Step 3: tert-butyl (2S,4S)-4-[(tert-butyldimethylsilyl)oxy]-2-[[(2-fluoro-4-iodopyridin-3yl)oxy]methyl]pyrrolidine-1-carboxylate.

To a stirred mixture of 2-fluoro-4-iodopyridin-3-ol (400 mg, 1.674 mmol, 1.0 equiv), tert-butyl (2S,4S)-4-[(tert-butyldimethylsilyl)oxy]-2-( hydroxymethyl)pyrrolidine-1-carboxylate (555 mg, 1.674 mmol, 1.00 equiv) and PPh ₃ (658 mg, 2.511 mmol, 1.50 equiv) in THF (4 ml) add DEAD (437 mg, 2.51 mmol , 1.50 equiv) dropwise at 0°C in a nitrogen atmosphere. The resulting mixture was stirred for 2 hours at room temperature under a nitrogen atmosphere, then quenched with water at 0°C and extracted with EtOAc. The combined organic layers are washed with water and brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue is purified by column chromatography on silica gel, eluted

- 43 044940

EtOAc/PE (0-30%) to give the title compound (735 mg, 79.4%).

Step 4: (3S,5S)-5-[[(2-fluoro-4-iodopyridin-3-yl)oxy]methyl]pyrrolidin-3-ol hydrochloride.

To a stirred solution of tert-butyl (2S,4S)-4-[(tert-butyldimethylsilyl)oxy]-2-[[(2-fluoro-4iodopyridin-3-yl)oxy]methyl]pyrrolidine-1-carboxylate (750 mg , 1.357 mmol, 1.0 equiv) add a solution of 3.0 M HCl in 1,4-dioxane (5 ml, 15 mmol, 11 equiv) dropwise at 0-5°C. After stirring at room temperature for 1 hour, the reaction mixture was concentrated under reduced pressure and the residue triturated with Et ₂ O. The precipitates were collected by filtration and dried under reduced pressure to obtain the title compound (400 mg, 78.7%).

Step 5: (6aS,8S)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazine-8- ol.

Mixture of (3S,5S)-5-[[(2-fluoro-4-iodopyridin-3-yl)oxy]methyl]pyrrolidin-3-ol hydrochloride (400 mg, 1.068 mmol, 1.00 equiv) and K2CO3 (443 mg, 3.205 mmol, 3.00 equiv) in EtOH (4 ml) was stirred for 2 hours at 60°C. After cooling to rt, the resulting mixture was diluted with water and extracted with EtOAc. The combined organic layers are washed with water and brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-40%) to give the title compound (300 mg, 88.3%).

Step 6: (6aS,8S)-4-iodo-8-methoxy-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4] oxazine

(6aS,8S)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1.2K stirred solution

d][1,4]oxazin-8-ol (100.00 mg, 0.314 mmol, 1.0 equiv) and Ag ₂ O (364.23 mg, 1.572 mmol, 5.0 equiv) in DMF (1.00 ml) add MeI (133.86 mg, 0.943 mmol, 3.00 equiv) dropwise at room temperature, and the resulting mixture is stirred for 4 hours at 50°C. After cooling to room temperature, the reaction mixture is filtered. The filtrate is washed with H ₂ O and brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with PE/EA (2/1) to give the title compound (89 mg, 85.2%).

Step 7: tert-butyl ((3S,4S)-8-(3-(hydroxymethyl)-5-(((6aS,8S)-8-methoxy-6a,7,8,9-tetrahydro-6Hpyrido[3, 2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4,5]decane- 4yl)carbamate.

(6aS,8S)-4-iodo-8-methoxy-6a,7,8,9-tetrahydro-6H-pyrido[3,2 mixtures

To stirred b]pyrrolo[1,2-d][1,4]oxazine (30.00 mg, 0.090 mmol, 1.0 equiv) and tert-butyl N-[(3S,4S)-8-[3 ( Hydroxymethyl)-5-sulfanylpyrazin-2-yl]-3-methyl-2-oxa-8-azaspiro[4,5]decan-4-yl]carbamate (37.08 mg, 0.090 mmol, 1.0 equiv) in dioxane (0.30 ml) add Pd ₂ (dba) ₃ (24.81 mg, 0.027 mmol, 0.30 equiv), Xantphos (15.68 mg, 0.027 mmol, 0.30 equiv) and DIEA (35.02 mg, 0.271 mmol, 3.0 equiv) at room temperature under N2 atmosphere. The resulting mixture was stirred for 1 hour at 80°C under N2 atmosphere.

After cooling to room temperature, the reaction mixture was concentrated in vacuo and the residue was purified by silica gel column chromatography, eluting with DCM/MeOH (10/1) to give the title compound (25 mg, 45.6%).

Step 8: (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)- 8-methoxy6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol formate

- 44 044940

TFA (1.3 ml) is added to the solution of tert-butyl ((3S,4S)-8-(3-(hydroxymethyl)-5-(((6aS,8S)-8methoxy-6a,7,8,9-tetrahydro -6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)-3methyl-2-oxa-8-azaspiro[4 ,5]decan-4-yl)carbamate (130.0 mg, 0.21 mmol, 1.0 equiv) in DCM (3.0 ml) at room temperature. After stirring at rt for 2 hours, the reaction solution was concentrated and the residue was purified by pre-HPLC to give the product (19 mg, 16.1%). MS (ER, m/z): [M+1] ⁺ = 515.3.

Example 6.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8R)-8- methoxy6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol.

Step 1: (6aS,8R)-4-iodo-8-methoxy-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4] oxazine

The compound (6aS,8R)-4-iodo-8-methoxy-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazine is synthesized the method described in example 5, steps 1-6 using 1-(tert-butyl) 2-methyl (2S,4S)-4-hydroxypyrrolidine-1,2-dicarboxylate in step 1.

Step 2: (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8R)- 8-methoxy6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol .

(3-((3S,4S)-4-Amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8R)-8-methoxy -6a,7,8,9tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol is synthesized by processing in a similar way the one described in example 5, stages 7-8. MS (ER, m/z): [M+1] ⁺ = 515.3.

Example 7.

Synthesis of (6aS,8S)-4-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-( Hydroxymethyl)pyrazin-2-yl)thio)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-8-ol.

The title compound was synthesized by the method described in Example 5, steps 7-8 using (6aS,8S)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[ 1,2-d][1,4]oxazin-8-ol at stage 7. MS (ER, m/z): [M+1]+ = 501.3.

Example 8.

Synthesis of (6aS,8R)-4-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-( Hydroxymethyl)pyrazin-2-yl)thio)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-8-ol.

- 45 044940

The title compound was synthesized by treatment similar to that described in Example 7. MS (ES, m/z): [M+1]+= 501.3.

Example 9.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aR,8R)-8- methoxy6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol.

The title compound was synthesized by workup similar to that described in Example 5, steps 2-8 using 1-tert-butyl 2-methyl (2R,4R)-4-methoxypyrrolidine-1,2dicarboxylate in step 2. MS (ES, m/z): [M+1]+= 515.3.

Example 10.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aR,8S)-8- methoxy6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol.

The title compound was synthesized by workup similar to that described in Example 5, Steps 2-8 using tert-butyl (2R,4S)-2-(hydroxymethyl)-4-methoxypyrrolidine-1carboxylate in Step 2. MS (ES, m/ z): [M+1]+ = 515.2.

Example 11.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)-8- (((S)-tetrahydrofuran-3-yl)oxy)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin- 4-yl)thio)pyrazin-2yl)methanol.

Step 1: (6aS,8S)-4-iodo-8-(((S)-tetrahydrofuran-3-yl)oxy)-6a,7,8,9-tetrahydro-6H-pyrido[3,2b]pyrrolo[ 1,2-d][1,4]oxazine.

To a stirred solution of (3R)-oxolan-3-yl 4-methylbenzenesulfonate (457 mg, 1.886 mmol, 3 equiv) and (6aS,8S)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[ 3,2-b]pyrrolo[1,2-d][1,4]oxazin-8-ol (200 mg, 0.629 mmol, 1.00 equiv) in DMF (2.00 ml) add NaH (88 mg, 2,200 mmol, 3.5 equiv, 60%) in portions at 0° C. under a nitrogen atmosphere, and the resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere. After cooling to 0°C, the reaction is quenched with water and the resulting mixture is diluted with water and extracted with EtOAc. The combined organic layers are washed with water and brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by pre-TLC (EtOAc 100%) to give the title compound (70 mg, 28.7%).

Step 2: (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)- 8-(((S)- 46 044940 tetrahydrofuran-3-yl)oxy)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d] [1, 4]oxazin-4-yl)thio)pyrazin-2-yl)methanol.

The title compound was synthesized from (6aS,8S)-4-iodo-8-(((S)-tetrahydrofuran-3yl)oxy)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b ]pyrrolo[1,2-d][1,4]oxazine by treatment similar to that described in example 5, steps 7-8. MS (ER, m/z): [M+1]+=571.3.

Example 12.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)-8- (((R)tetrahydrofuran-3-yl)oxy)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4yl )thio)pyrazin-2-yl)methanol.

The title compound was synthesized from (38)-oxolan-3-yl 4-methylbenzenesulfonate by treatment similar to that described in Example 11. MS (ES, m/z): [M+1] ⁺ =571.3.

Example 13.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azacniro[4,5]decan-8-yl)-6-(((6aS,8S)-8- (oxetan-3yloxy)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2 -yl)methanol.

Step 1: (6aS,8R)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]nirrolo[1,2-d][1,4]oxazin-8yl trifluoromethanesulfonate .

To a stirred solution of (6aS,8R)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]-pyrrolo[1,2d][1,4]oxazin-8-ol (400 mg, 1.257 mmol, 1.00 equiv) and DIEA (244 mg, 1.886 mmol, 1.50 equiv) in DCM (4.00 ml) add Tf ₂ O (426 mg, 1.509 mmol, 1.20 equiv) drop by drop at 5°C. The reaction solution is stirred at 5°C for 30 minutes and then concentrated in vacuo. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-40%) to give the title compound (300 mg, 53.0%) as a white solid.

Step 2: (6aS,8S)-4-iodo-8-(oxetan-3-yloxy)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2d][ 1,4]oxazine.

To a stirred solution of oxetan-3-ol (99 mg, 1.336 mmol, 2.00 equiv) in DMF (3.00 ml) add 60% NaH (67 mg, 1.67 mmol, 2.50 equiv) at 5°C . The mixture is stirred at this temperature for 1 hour. (6aS,8R)-4-Iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1 ,4]oxazin-8-yl trifluoromethanesulfonate (300.00 mg, 0.666 mmol, 1.00 equiv) was added at 20°C. The resulting mixture is heated to 50°C and stirred for 10 minutes. The reaction is quenched with water at 5°C and the resulting mixture is extracted with EtOAc. The combined organic layers are washed with water and brine and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-100%) to give the title compound (20 mg, 8.0%) as a white solid.

- 47 044940

Step 3: (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)- 8-(oxetan-3yloxy)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazine -2-yl)methanol.

The title compound was synthesized by workup similar to that described in Example 5, steps 7-8 using (6aS,8S)-4-iodo-8-(oxetan-3-yloxy)-6a,7,8,9-tetrahydro -6Hpyrudo[3,2-b]pyrrolo[1,2-d][1,4]oxazine at stage 7. MS (ER, m/z): [M+1]+=557.3.

Example 14.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)-8- (methoxymethoxy)6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl) methanol.

Step 1: (6aS,8S)-4-iodo-8-(methoxymethoxy)-6a,7,8,9-tetrahydro-6H-nirido[3,2-b]pyrrolo[1,2d][1,4] oxazine

To a stirred solution of (6aS,8S)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazine-8- ol (60 mg, 0.189 mmol, 1.00 equiv) and DIEA (49 mg, 0.377 mmol, 2.00 equiv) in DME (1.00 ml) add bromo(methoxy)methane (35.35 mg, 0.283 mmol, 1.50 equiv) dropwise at room temperature under N2 atmosphere. The resulting mixture was stirred for 4 hours at room temperature under N2 and then quenched with H ₂ O. The resulting mixture was extracted with EA and the combined organic layers were dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE/EA (1/1) to give the title compound (50 mg, 73.2%) as a light yellow solid.

Step 2: (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)- 8(methoxymethoxy)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d] [1,4]oxazin-4-yl)thio)pyrazin-2yl) methanol

The title compound was synthesized by workup similar to that described in Example 5, steps 7-8 using tert-butyl ((3S,4S)-8-(3-(hydroxymethyl)-5-(((6aS,8S)- 8(methoxymethoxy)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-ul)thio)pyrazin-2yl) -3-methyl-2-oxa-8-azaspiro[4,5]decan-4-yl)carbamate at step 7. MS (ES, m/z): [M+1]+=545.2.

Example 15.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)-8- (2-methoxyethoxy)6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2- yl)methanol.

The title compound is synthesized by processing similar to that described in

- 48 044940 example 14 using 2-bromoethyl methyl ether in step 1. MS (ES, m/z): [M+1]+=559.3.

Example 16.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)-8- (cyclopropylmethoxy)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl )methanol.

Ό

The title compound was synthesized by workup similar to that described in Example 14 using (bromomethyl)cyclopropane r in step 1. MS (ES, m/z): [M+1]+=555.4.

Example 17.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((S)-6a',7 '-dihydro-6Ή,9Ήspiro [cyclopropan-1, 8'-pyrido[3,2-b]pyrrolo [1,2-d][1,4]oxazin] -4'-yl)thio)pyrazin-2- yl)methanol.

_H2N \

Step 1: (S)-4-iodo-6a',7-dihydro-6Ή,9Ή-spiro[cyclopropane-1,8'-pyrido[3,2-b]pyrrolo[1,2-d][1, 4]oxazine].

Compound (S)-4'-iodo-6a',7'-dihydro-6Ή,9Ή-spiro[cyclopropane-1,8'-nirido[3,2-b]pyrrolo[1,2d][1,4] oxazine] is synthesized by processing similar to that described in example 1, steps 1-4 using 5-tert-butyl 6-methyl (6S)-5-azaspiro[2,4]heptane-5,6-dicarboxylate in step 1.

Step 2: (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((S)-6a' ,7'-dihydro6Ή,9Ή-spiro[cyclopropan-1,8'-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin]-4'-yl)thio)pyrazin- 2yl)methanol.

The title compound is synthesized by processing similar to that described in Example 1, steps 5-8. MS (ER, m/z): [M+1]+=511.3.

Example 18 and 19.

Synthesis of (6aS,8S)-4-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decane-8-ul)-6-( Hydroxymethyl)pyrazin-2-yl)thio)-8-methyl-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]-pyrrolo[1,2-d][1,4]oxazine -8-ol and (6aS,8R)-4-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl) -6-(hydroxymethyl)pyrazin-2yl)thio)-8-methyl-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4] oxazin-8-ol.

The stereochemistry of the tertiary alcohol is arbitrarily specified in 18 and 19.

Step 1: (S)-4-iodo-6a,7-dihydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-8(9H)-one.

- 49 044940

To a stirred solution of oxalyl chloride (209.47 mg, 1.650 mmol, 1.5 equiv) in DCM (1.4 ml) add DMSO (257.89 mg, 3.301 mmol, 3.0 equiv) in DCM (0.2 ml) at drops at -78°C under a nitrogen atmosphere and stirred at this temperature for 30 minutes. A solution of (6aS)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazine-8 is added to the solution obtained above -ol (350.00 mg, 1.100 mmol, 1.00 equiv) in DCM (2.0 ml) at -78°C. After stirring at -78°C for 30 minutes, DIEA (853.1 mg, 6.6 mmol, 6.0 equiv) was added dropwise at -78°C. The resulting mixture was stirred for an additional 30 minutes at -78°C, then warmed to room temperature over 30 minutes. After stirring at rt for 10 minutes, the reaction mixture is cooled to 5°C and quenched by adding sat. NH ₄ Cl aq. solution at 5°C. The resulting mixture was extracted with EtOAc and the combined organic layers were washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated to give the title compound (250 mg, 71.8%).

Step 2: 2-ethylhexyl 3-(((S)-8-oxo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4 ]oxazin-4-yl)thio)propanoate.

Solution of (S)-4-iodo-6a,7-dihydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-8(9H)-one (220.00 mg, 0.696 mmol, 1.00 equiv), 2-ethylhexyl 3-sulfanylpropanoate (182.37 mg, 0.835 mmol, 1.20 equiv), Pd ₂ (dba) ₃ (63.73 mg, 0.070 mmol, 0.10 equiv), Xantphos (40.27 mg, 0.070 mmol, 0.10 equiv) and DIEA (269.85 mg, 2.088 mmol, 3.00 equiv) in dioxane (4.40 ml) stir for 1 hour at 100° C in a nitrogen atmosphere. The resulting mixture was concentrated in vacuo and the residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-50%), to give the title compound (280 mg, 99%).

Step 3: 2-ethylhexyl 3-(((6aS)-8-hydroxy-8-methyl-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d] [1,4]oxazin-4-yl)thio)propanoate.

To a stirred solution of 2-ethylhexyl 3-(((S)-8-oxo-6a,7,8,9-tetrahydro-6H-pyrido[3,2b]pyrrolo[1,2-d][1,4]oxazine -4-yl)thio)propanoate (160 mg, 0.394 mmol, 1.00 equiv) in THF (1.60 ml) add 2.0 M bromo(methyl)magnesium (0.24 ml, 0.48 mmol, 1 .22 equiv) dropwise at 5°C under a nitrogen atmosphere, and the resulting mixture was stirred at 5°C for 1 hour. The reaction was quenched by adding sat. NH ₄ Cl aq. solution at 5°C and extracted with EtOAc. The combined organic layers are washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-100%) to give the title compound (90 mg, 54.1%).

Step 4: (6aS,8S)-4-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6 -(hydroxymethyl)pyrazin-2-yl)thio)-8-methyl-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]-pyrrolo[1,2-d][1,4 ]oxazin-8-ol and (6aS,8R)-4-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decane-8- yl)-6-(hydroxymethyl)pyrazin-2yl)thio)-8-methyl-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1, 4]oxazin-8-ol.

The title compounds are synthesized from 2-ethylhexyl 3-(((6aS)-8-hydroxy-8-methyl6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d ][1,4]oxazin-4-yl)thio)propanoate by treatment similar to that described in example 1, steps 6-8. MS (ER, m/z): [M+1]+=515.3.

Example 20.

Synthesis of (6aS,8S)-4-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-( Hydroxymethyl)pyrazin-2-yl)thio)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazine-8-carbonitrile.

- 50 044940

Step 1: (6a8,8K)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-b][1,4]oxazine-8- silt methanesulfonate.

To a stirred solution of (6a8,8K)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2d][1,4]oxazin-8-ol ( 400.00 mg, 1.257 mmol, 1.00 equiv) in DCM (6.00 ml) add TEA (190.86 mg, 1.886 mmol, 1.50 equiv) and MsCl (172.85 mg, 1.509 mmol, 1.00 equiv) 20 equiv) at room temperature. After stirring for 16 hours at room temperature, the reaction mixture is cooled to 0°C and quenched with water. The resulting mixture was extracted with EtOAc and the combined organic layers were washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-50%) to give the title compound (450 mg, 90.3%).

Step 2: (6aS,8S)-4-iodo-6a,7,8,9-τeτpahydro-6H-πpyrido[3,2-b]πpyrrolo[l,2-d][l,4]ocasin-8carbonitrile.

To a stirred solution of (6a8,8K)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2d][1,4]oxazin-8-yl methanesulfonate (300.00 mg, 0.757 mmol, 1.00 equiv) in DMF (5.00 ml) add NaCN (55.66 mg, 1.136 mmol, 1.50 equiv) at room temperature. After stirring at 75°C for 16 hours, the reaction mixture is cooled at room temperature and quenched with water. The mixture was then extracted with EtOAc and the combined organic layers were washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-50%) to give the title compound (140 mg, 56.5%).

Step 3: (6a8.88)-4-((5-((38.48)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6 (hydroxymethyl)pyrazin-2-yl)thio)-6a,7,8,9-tepahidro-6H-pyrido[3,2-b]pyrrolo[l,2-d][l,4]-oxazine-8carbonitrile.

The compound is synthesized from (6a8,88)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2d][1,4]oxazine-8-carbonitrile by treatment , similar to that described in example 5, stages 7-8. [M+1] ⁺ =510.2.

Example 21.

Synthesis of (3-((38.48)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(((6a8.88)-8( methylsulfonyl)-6a,7,8,9-τeτpahydro-6H-pyrido[3,2-b]πppolo[l,2-d][l,4]ocasin-4-yl)thio)pyrazin-2yl)methanol.

Step 1: 1-tert-butyl 2-methyl (28.4K)-4-(methanesulfonyloxy)pyrrolidine-1,2-dicarboxylate.

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To a stirred solution of 1-tert-butyl 2-methyl (2S,4R)-4-hydroxypyrrolidine-1,2dicarboxylate (9 g, 36.7 mmol, 1.0 equiv) and TEA (7.43 g, 73.4 mmol , 2.0 equiv) in DCM (100 ml) add MsCl (5 g, 43.6 mmol, 1.2 equiv) dropwise at 0°C, and the resulting mixture is stirred for one hour at 0°C. The reaction is quenched with water and extracted with DCM. The combined organic layers are washed with water and brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-50%) to give the title compound (10.5 g, 88.5%).

Step 2: 1-(tert-butyl)2-methyl (2R,4R)-4-methoxypyrrolidine-1,2-dicarboxylate.

-tert-butyl 2-methyl (2S,4R)-4-(methanesulfonyloxy)pyrrolidine-1,2-dicarboxylate

To a solution (4.30 g, 13.298 mmol, 1.00 equiv) in dry DMF (20.00 ml) add NaSCH ₃ (1.07 g, 15.27 mmol,

1.07 equiv) at 30°C. After stirring overnight at rt, the reaction mixture was poured into water and extracted with EtOAc. The combined organic layers are washed with water and brine, dried over Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with PE/EA (1/1) to give the title compound (4 g, 109.2%).

Step 3: tert-butyl (2S,4S)-2-(hydroxymethyl)-4-(methylthio)pyrrolidine-1-carboxylate.

To a stirred mixture of 1-tert-butyl 2-methyl (2S,4S)-4-(methylsulfanyl)pyrrolidine-1,2dicarboxylate (3.70 g, 13.44 mmol, 1.0 equiv) in THF (37.0 ml ) add LiAlH ₄ (0.76 g, 20.03 mmol, .5 equiv) in portions at 0°C. After stirring at room temperature for 2 hours, the reaction mixture is quenched with Na ₂ SO ₄ -10 H ₂ O at room temperature. The resulting mixture was filtered and the filtrate was concentrated under reduced pressure to give the title compound (2.6 g, 78.2%).

Step 4: tert-butyl (2S,4S)-2-(hydroxymethyl)-4-(methylsulfonyl)pyrrolidine-1-carboxylate.

tert-butyl (2S,4S)-2-(hydroxymethyl)-4-(methylsulfanyl) To a stirred solution of pyrrolidine-1-carboxylate (3.70 g, 14.959 mmol, 1.00 equiv) in DCM (40 ml) at room temperature temperature add m-CPBA (77%, 12.91 g, 57.60 mmol, 3.85 equiv) in portions over 5 hours. The resulting mixture is diluted with DCM and washed with sat. aq. NaHCO ₃ solution and brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with DCM/MeOH (20/1) to give the title compound (1 g, 23.9%).

Step 5: (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)- 8(methylsulfonyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2yl) methanol

The title compound was synthesized from tert-butyl (2S,4S)-2-(hydroxymethyl)-4(methylsulfonyl)pyrrolidine-1-carboxylate by the method described in Example 1, Steps 2-8. MS (ER, m/z): [M+1]+=563.3.

Example 22

Synthesis of (6aS,8S)-4-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-( hydroxymethyl- 52 044940 yl)pyrazin-2-yl)thio)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d]-[1,4]oxazine -8-yl carbamate

Step 1: (6aS,8S)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazine-8- silt carbamate.

To a stirred solution of (6aS,8S)-4-iodo-6α,7,8,9-tetraαhydro-6H-pyrido[3,2-b]pyrrolo[1,2d][1,4]oxazin-8-ol ( 150.0 mg, 0.472 mmol, 1.0 equiv) to DCM (1.80 ml), add trichloroethanecarbonyl isocyanate (222.08 mg, 1.179 mmol, 2.5 equiv) dropwise at 5°C, and the resulting mixture is stirred at at this temperature for approximately 30 minutes. To the above solution, K ₂ CO ₃ (129.88 mg, 0.940 mmol, 2.0 equiv) and MeOH (1.80 ml) were added at room temperature, and the resulting mixture was stirred for an additional 6 hours at room temperature. The mixture is diluted with water and extracted with EtOAc. The combined organic layers are washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-100%) to give the title compound (100 mg, 58.7%).

Step 2: (6aS,8S)-4-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6 -(hydroxymethyl)pyrazin-2-yl)thio)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]-oxazine-8 -yl carbamate.

The title compound was synthesized from tert-butyl (2S,4S)-2-(hydroxymethyl)-4(methylsulfonyl)pyrrolidine-1-carboxylate by treatment similar to that described in Example 5, Steps 7-8. MS (ER, m/z): [M+1]+=544.3.

Example 23

Synthesis of (6aS,8R)-4-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-( Hydroxymethyl)pyrazin-2-yl)thio)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-8-yl carbamate .

The title compound is synthesized from ((6aS,8R)-4-iodo-6a,7,8,9-tetrahydro-6Hpyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin- 8-ol as described in example 22. MS (ER, m/z): [M+1]+=544.2.

Example 24.

Synthesis of (6-(((6aS,8S)-8-(1H-pyrazol-1-yl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]-pyrrolo[1,2d ][1,4]oxazin-4-yl)thio)-3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl) pyrazin-2yl)methanol.

Step 1: (6aS,8S)-4-iodo-8-(III-pyrazol-1-yl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]-pyrrolo[1, 2d][1,4]oxazine.

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To a stirred solution of pyrazole (17.18 mg, 0.252 mmol, 1.0 equiv) in DMF (0.75 ml) add NaH (20.19 mg, 0.505 mmol, 2.0 equiv, 60%) in portions at 5°C , and the resulting mixture was stirred at 5°C for 1 hour. (6aS,8R)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2b]pyrrolo[1, 2-d][1,4]oxazin-8-yl methanesulfonate (100.00 mg, 0.252 mmol, 1.00 equiv) at room temperature and the resulting mixture was stirred at 50°C. After cooling to 0°C, the reaction is quenched by adding water and then extracted with EtOAc. The combined organic layers are washed with water, brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-100%) to give the title compound (60 mg, 64.5%).

Step 2: (6-(((6aS,8S)-8-(1H-pyrazol-1-yl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1, 2d][1,4]oxazin-4-yl)thio)-3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl )pyrazin-2yl)methanol.

The title compound is synthesized from (6aS,8S)-4-iodo-8-(1H-pyrazol-1-yl)-6a,7,8,9tetrahydro-6H-pyrido[3,2-b]pyrrolo[1, 2-d][1,4]oxazine by treatment as described in Example 5, steps 7-8. MS (ER, m/z): [M+1] ⁺ =551.3.

Example 25.

Synthesis of (6-(((6aS,8R)-8-(1H-pyrazol-1-yl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]-pyrrolo[1,2d ][1,4]oxazin-4-yl)thio)-3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl) pyrazin-2yl)methanol.

The title compound is synthesized from (6aS,8S)-4-iodo-6a,7,8,9-tetrahydro-6Hpyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazine-8 -yl methanesulfonate by the method described in example 24. MS (ES, m/z): [M+1] ⁺ =551.3.

Example 26 and 27.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((R)-6a,7, 9,10-tetrahydro6H-[1,4]oxazino[4,3-d]pyrido[3,2-b][1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol and (3 -((3S,4S)-4-amino3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((S)-6a,7,9,10-tetrahydro -6H-[1,4]oxazino[4,3d]pyrido[3,2-b][1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol.

The title compounds were synthesized by workup similar to that described in Example 5, steps 2-8 using tert-butyl 3-(hydroxymethyl)morpholine-4-carboxylate in step 2. MS (ES, m/z): [M +1]+=501.2.

Example 28.

Synthesis of 4-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-αzaspiro[4,5]decan-8-yl)-6-(hydroxymethyl)pyrazin-2yl) thio)-6a,7,9,10-tetrahydro-6H-pyrido[3,2-b][1,4]thiazino[4,3-d][1,4]oxazine 8,8-dioxide.

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Step 1: 4-(tert-butoxycarbonyl)thiomorpholine-3-carboxylic acid.

To a stirred solution of thiomorpholine-3-carboxylic acid (3000 mg, 20.382 mmol, 1.00 equiv) and Et ₃ N (4125 mg, 40.764 mmol, 2.00 equiv) in DCM (50.00 ml) add Boc ₂ O ( 6672 mg, 30.57 mmol, 1.50 equiv) in portions at room temperature under N2. The resulting mixture was stirred for 16 hours at room temperature under N2 and then diluted with water and extracted with DCM. The combined organic layers are dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with DCM/MeOH (10/1) to give the title compound (3.2 g, 63.5%) as a light yellow oil.

Step 2: tert-butyl 3-(hydroxymethyl)thiomorpholine-4-carboxylate.

To a stirred solution of 4-(tert-butoxycarbonyl)thiomorpholine-3-carboxylic acid (2000 mg, 8.087 mmol, 1.00 equiv) in THF (20 ml) add 1 M BH3-THF (16.18 ml, 16.180 mmol, 2. 00 equiv) dropwise at 0°C in an N2 atmosphere. The resulting mixture was stirred for 16 hours at 45°C in an atmosphere

N2 and then quenched with MeOH at 0°C. The resulting mixture was concentrated in vacuo and the residue was purified by silica gel column chromatography, eluting with DCM/MeOH (10/1) to give the title compound (1.8 g, 95.4%) as a light yellow oil.

Step 3: tert-butyl 3-(hydroxymethyl)thiomorpholine-4-carboxylate 1,1-dioxide.

A solution of tert-butyl 3-(hydroxymethyl)thiomorpholine-4-carboxylate (500 mg, 2.143 mmol, 1.00 equiv) and m-CPBA (77%, 851 mg, 3.78 mmol, 1.77 equiv) in DCM ( 10 ml) was stirred for 16 hours at room temperature under N2 atmosphere. The reaction is quenched with saturated NaHCO ₃ (5 ml) at room temperature. The resulting mixture was extracted with DCM. The combined organic layers are washed with saturated NaHCO3, dried over anhydrous Na2SO4. After filtration, the filtrate is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with DCM/MeOH (10/1) to give the title compound (400 mg, 70.4%) as a light yellow oil.

Step 4: 3-(hydroxymethyl)thiomorpholine 1,1-dioxide; trifluoroacetic acid.

To a stirred solution of tert-butyl 3-(hydroxymethyl)thiomorpholine-4-carboxylate 1,1 dioxide (180 mg, 0.678 mmol, 1.00 equiv) in DCM (5.0 ml) add TFA (3.868 g, 33.921 mmol, 50. 00 equiv) at room temperature. The resulting mixture was stirred for 2 hours at room temperature. The resulting mixture was concentrated under reduced pressure to obtain the title compound (170 mg, 89.74%) as a white solid.

Step 5: 4-iodo-6a,7,9,10-tetrahydro-6H-pyrido[3,2-b][1,4]thiazino[4,3-d][1,4]oxazine 8,8- dioxide

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To a stirred solution of 3-(hydroxymethyl)thiomorpholine 1,1-trifluoroacetic acid dioxide (180 mg, 0.645 mmol, 1.00 equiv) and 2-fluoro-4-iodopyridin-3-ol (154 mg, 0.645 mmol, 1.00 equiv) in toluene (3.0 ml) add PPh ₃ (254 mg, 0.967 mmol, 1.50 equiv) and DEAD (168 mg, 0.967 mmol, 1.50 equiv) at room temperature under nitrogen. The resulting mixture was stirred for 16 hours at 60°C under a nitrogen atmosphere and then concentrated under reduced pressure. The residue was purified by pre-TLC (PE/EtOAc 5:1) to give the title compound (45 mg, 19.1%) as a light yellow solid.

Step 6: 4-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(hydroxymethyl)pyrazin- 2-yl)thio)-6a,7,9,10-tetrahydro-6H-pyrido[3,2-b][1,4]thiazino[4,3-d][1,4]oxazine 8,8- dioxide

The title compounds were synthesized by a similar treatment to that described in Example 5, Steps 7-8 using 4-iodo-6a,7,9,10-tetrahydro-6H-pyrido[3,2-b][1,4 ]thiazino[4,3d][1,4]oxazine 8,8-dioxide at step 7. MS (ER, m/z): [M+1]+=549.2.

Example 29 and 30.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-αzaspiro[4,5]decan-8-yl)-6-(((6aS,8S)-8- (methoxymethyl)6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl) methanol [29] and (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8R )-8-(methoxymethyl)-6a,7,8,9tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2 -yl)methanol [30].

Step 1: 1-(tert-butyl)2-methyl (S)-4-(methoxymethylene)pyrrolidine-1,2-dicarboxylate.

To a stirred mixture of 1-(tert-butyl) 2-methyl (S)-4-oxopyrrolidine-1,2-dicarboxylate (4.00 g, 16.443 mmol, 1.00 equiv) and K ₂ CO ₃ (1.50 g , 10.853 mmol, 0.66 equiv) to MeOH (80.0 ml) add dimethyl (1-diazo-2-oxopropyl)phosphonate (9.48 g, 49.330 mmol, 3.00 equiv) at 0°C under nitrogen atmosphere . After stirring for 12 hours at room temperature, the reaction mixture was poured into water and the resulting mixture was extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with PE/EtOAc (3:1) to give the title compound (1.45 g, 32.5%) as a light yellow oil.

Step 2: 1-(tert-butyl)2-methyl (2S)-4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate.

To a stirred solution of 1-(tert-butyl)2-methyl (S)-4-(methoxymethylene)pyrrolidine-1,2dicarboxylate (1.45 g, 5.344 mmol, 1.00 equiv) and 10% Pd/C (145 mg ) to MeOH (20.0 mL) add MgO (220 mg, 5.46 mmol, 1.02 equiv) at room temperature under hydrogen atmosphere. After stirring for 2 hours at room temperature, the resulting mixture was filtered. The filtrate was concentrated under reduced pressure to give the title compound (1.35 g, 92.42%) as a light yellow oil, which was used immediately in the next step without further purification.

Step 3: tert-butyl (2S)-2-(hydroxymethyl)-4-(methoxymethyl)pyrrolidine-1-carboxylate.

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To a stirred solution of 1-(tert-butyl)2-methyl (2S)-4-(methoxymethyl)pyrrolidine-1,2dicarboxylate (180 mg, 0.659 mmol, 1.00 equiv) in THF (4.0 mL) add LiAlH ₄ (37 mg, 0.975 mmol, 1.50 equiv) at room temperature. After stirring for 2 hours at room temperature, the resulting mixture is diluted with DCM and quenched with water at 0°C. After filtration, the filter cake is washed with DCM. The organic layer was washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with PE/EtOAc (1:1) to give the title compound (90 mg, 55.7%) as a light yellow oil.

Step 4: tert-butyl pyrrolidine-1-carboxylate.

(2S)-2-(((2-fluoro-4-iodopyridin-3-yl)okCi)methyl)-4-(methokCimethyl)-

To a stirred solution of tert-butyl (2S)-2-(hydroxymethyl)-4-(methoxymethyl)pyrrolidin-1-carboxylate (90 mg, 0.367 mmol, 1.00 equiv) and 2-fluoro-4-iodopyridin-3-ol (88 mg, 0.367 mmol, 1.00 equiv) in THF (2.00 ml) add PPh ₃ (144 mg, 0.550 mmol, 1.50 equiv) and DEAD (96 mg, 0.55 mmol, 1.50 equiv) at room temperature in a nitrogen atmosphere. After stirring for 16 hours at room temperature, the reaction mixture is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-60%) to give the title compound (115 mg, 67.2%) as a light yellow oil.

Step 5: 2-fluoro-4-iodo-3-(((2S)-4-(methoxymethyl)pyrrolidin-2-yl)methoxy)pyridine hydrogen chloride.

To a stirred solution of tert-butyl (2S)-2-(((2-fluoro-4-iodopyridin-3-yl)oxy)methyl)-4(methoxymethyl)pyrrolidine-1-carboxylate (115 mg, 0.247 mmol, 1. 00 equiv) in dioxane (1.0 ml) add HCl (4.0 M in dioxane, 0.62 ml, 2.48 mmol, 10.00 equiv) at room temperature. After stirring for 5 hours at room temperature, the reaction mixture was concentrated under reduced pressure to obtain the title compound (110 mg, crude) as a light yellow solid, which was used immediately in the next step without further purification. MS (ER, m/z): [M+1]+=367.0.

Step 6: d][1,4]oxazine.

(6aS)-4-iodo-8-(methokCymethyl)-6a,7,8,9-tetraαhydro-6H-pyrido[3,2-b]nirrolo[1,2

a solution of 2-fluoro-4-iodo-3-(((2S,4S)-4-(methoxymethyl)pyrrolidin-2-yl)to stirred methoxy)pyridine hydrogen chloride (110 mg, 0.273 mmol, 1.00 equiv) in EtOH (3.0 ml) add K ₂ CO ₃ (208 mg, 1.50 mmol, 5.51 equiv) at room temperature. After stirring for 2 hours at 60°C, the reaction mixture is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-60%) to give the title compound (80 mg, 2 steps, 93.5%) as a light yellow oil.

Step 7: tert-butyl ((3S,4S)-8-(3-(hydroxymethyl)-5-(((6aS)-8-(methoxymethyl)-6a,7,8,9-tetrahydro-6Hpyrido[3, 2-b]pyrrolo[1,2-d] [1,4]oxazin-4-yl)thio)pyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4,5]decane- 4yl)carbamate.

To a stirred mixture of (6aS)-4-iodo-8-(methokCimethyl)-6a,7,8,9-tetrahydro-6H-pyrido-[3,2b]pyrrolo[1,2-d][1,4]oxazine (80 mg, 0.231 mmol, 1.00 equiv), 5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3methyl-2-oxa-8-azaspiro[4,5]decane-8 -yl)-6-(hydroxymethyl)pyrazine-2-thiolate sodium (100 mg, 0.231 mmol,

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1.0 equiv), Pd ₂ (dba) ₃ (63 mg, 0.069 mmol, 0.30 equiv) and Xantphos (40 mg, 0.069 mmol, 0.30 equiv) in dioxane (1.50 ml) add DIEA (90 mg, 0.693 mmol, 3.00 equiv) at room temperature under nitrogen atmosphere. After stirring for 1 hour at 80°C, the reaction mixture is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with MeOH/DCM (0-10%) to give the title compound (110 mg, 75.7%) as a yellow solid.

Step 8: (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)- 8-(methoxymethyl)6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)-pyrazin-2 -yl)methanol [29] and (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-((( 6aS,8R)-8-(methoxymethyl)-6a,7,8,9tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio) pyrazin-2-yl)methanol [30].

To a stirred solution tert-butyl ((3S,4S)-8-(3-(hydroxymethyl)-5-(((6aS,8S)-8(methoxymethyl)-6a,7,8,9-tetrahydro-6H-pyrido [3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)-pyrazin-2yl)-3-methyl-2-oxa-8-azaspiro[4,5] Decan-4-yl)carbamate (70.00 mg, 0.111 mmol, 1.00 equiv) in DCM (1.0 ml) add TFA (0.20 ml, 2.61 mmol, 23.5 equiv) at room temperature . After stirring for 2 hours at room temperature, the reaction mixture is concentrated under reduced pressure. The mixture was purified by pre-HPLC to obtain 50 mg of crude product.

The crude product (50 mg) was then purified by chiral HPLC to give the title compound [29] (9.9 mg, 16.8%), MS (ES, m/z): [M+1]+=529.3 and [30] (5.5 mg, 9.4%), MS (ER, m/z): [M+1] ⁺ =529.3; as a white solid.

Example 31.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)-8- ((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin- 2yl)methanol.

Step 1: tert-butyl (2S,4S)-2-(hydroxymethyl)-4-(methoxymethyl)pyrrolidine-1-carboxylate.

To a stirred solution of (2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2carboxylic acid (180 g, 694.171 mmol, 1.00 equiv) in THF (1.8 L) add BH ₃ - Me ₂ S (173.00 ml, 1824.073 mmol, 2.63 equiv) dropwise at 0°C under N2. The resulting mixture was stirred for 16 hours at room temperature under N2 atmosphere. The reaction is quenched with MeOH (200 ml) at 0° C. and the resulting mixture is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EA (1/1) to give the title compound (160 g, 94%) as a colorless oil.

Step 2: tert-butyl (2S,4S)-2-(((2-fluoro-4-iodopyridin-3-yl)oxy)methyl)-4-(methoxymethyl)pyrrolidine-1-carboxylate.

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To a stirred solution of tert-butyl (2S,4S)-2-(hydroxymethyl)-4-(methoxymethyl)-pyrrolidine 1-carboxylate (7.0 g, 28.534 mmol, 1.00 equiv), 2-fluoro-4-iodopyridine- 3-ol (6.8 g, 28.534 mmol, 1.00 equiv) and PPh ₃ (11.2 g, 42.7 mmol, 1.50 equiv) in THF (140.00 ml) add DEAD (7.5 g, 43 mmol, 1.5 equiv) dropwise at rt under N2, and the resulting mixture was stirred for 16 h at room temperature. The reaction mixture was then concentrated under reduced pressure and the residue was purified by silica gel column chromatography, eluting with PE/EA (5/1) to give the title compound (11.5 g, 86.4%) as a light yellow oil.

Step 3: 2-fluoro-4-iodo-3-(((2S,4S)-4-(methoxymethyl)pyrrolidin-2-yl)methoxy)pyridine dihydrochloride.

Solution of tert-butyl (2S,4S)-2-(((2-fluoro-4-iodopyridin-3-yl)oxy)methyl)-4-(methoxymethyl)pyrrolidine-1-carboxylate (11.5 g, 24.663 mmol , 1.00 equiv) in 1,4-dioxane (20.0 ml) and 4.0 M HCl in 1,4-dioxane (50.0 ml) were stirred for 3 hours at room temperature under N2 atmosphere. The resulting mixture was concentrated under reduced pressure to give the title compound (14 g, crude) as a light yellow oil, which was used in the next step without further purification.

Step 4: (6aS,8S)-4-iodo-8-(methokCimethyl)-6α,7,8,9-tetrahydro-6H-nirido[3,2-b]nirrolo[1,2d][1,4] oxazine

Mixture of crude 2-fluoro-4-iodo-3-(((2S,4S)-4-(methoxymethyl)pyrrolidin-2-yl)-methoxy)pyridine dihydrochloride (14.0 g, 31.88 mmol, 1.00 equiv) and K ₂ CO ₃ (15.9 g, 115 mmol, 3.6 equiv) in EtOH (280.0 ml) were stirred for 3 hours at 60°C. After cooling to rt, the resulting mixture is filtered and the filter cake is washed with EtOAc. The combined filtrate is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE/EA (2/1) to give the title compound (6.6 g, 2 steps, 77%) as a light yellow solid.

Step 5: ((6aS,8S)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazine-8 -yl) methanol.

stirred mixture of (6aS,8S)-4-iodo-8-(methoxymethyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2b]pyrrolo[1,2-d][1,4]oxazine (250 mg, 0.722 mmol, 1.00 equiv) and NaI (357 mg, 2.383 mmol, 3.3 equiv) in CH3CN (5.00 ml) add SiCl ₄ (405 mg, 2.383 mmol, 3.30 equiv) at drops at room temperature in a nitrogen atmosphere. The resulting mixture was stirred for 2 hours at 80°C under a nitrogen atmosphere. The mixture is cooled to room temperature, diluted with water and the pH of the solution is adjusted to pH=8 with a saturated aqueous solution of NaHCO ₃ . The resulting mixture was extracted with DCM, and the organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated. The residue was purified by silica gel column chromatography, eluting with PE/EtOAc (1:1) to give the title compound (200 mg, 83.4%) as a yellow solid.

Step 6: (6aS,8S)-4-iodo-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2

b][1,4]oxazine.

To a stirred solution of ((6aS,8S)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2

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d][1,4]oxazin-8-yl)methanol (75 mg, 0.226 mmol, 1.00 equiv) and DIEA (88 mg, 0.68 mmol, 3.0 equiv) in

DCM (3.75 ml) add bromo(methoxy)methane (85 mg, 0.68 mmol, 3.0 equiv) dropwise at room temperature under nitrogen. The resulting mixture was stirred for 24 hours at room temperature under a nitrogen atmosphere. The reaction solution was purified by silica gel column chromatography, eluted with PE/EtOAc (1:1) to obtain the title compound (65 mg, 76.5%) as a yellow solid.

Step 7: tert-butyl ((3S,4S)-8-(3-(hydroxymethyl)-5-(((6aS,8S)-8-((methoxymethoxy)methyl)6a,7,8,9-tetrahydro- 6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)-3-methyl-2-oxa8-azaspiro[4, 5]decan-4-yl)carbamate.

To a stirred mixture of 5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8azaspiro[4,5]decan-8-yl)-6-(hydroxymethyl)pyrazine -Sodium -2-thiolate (55 mg, 0.127 mmol, 1.00 equiv), (6aS,8S)-4-iodo-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido [3,2-b]pyrrolo[1,2-d][1,4]oxazine (48 mg, 0.128 mmol, 1 equiv) and Xantphos (22 mg, 0.038 mmol, 0.30 equiv) in dioxane (2, 0 ml) add DIEA (82 mg, 0.63 mmol, 5 equiv) and Pd ₂ (dba) ₃ (35 mg, 0.038 mmol, 0.30 equiv) in portions at room temperature under nitrogen. The resulting mixture was stirred for 1 hour at 80°C under a nitrogen atmosphere and then diluted with water and extracted with EtOAc. The organic layer is dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with DCM/MeOH (10:1) to give the title compound (45 mg, 53.8%) as a brown solid.

Step 8: (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-αsniro[4,5]decan-8-yl)-6-(((6aS,8S)- 8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio) pyrazin-2yl)methanol.

To the stirred solution tert-butyl ((3S,4S)-8-(3-(hydroxymethyl)-5-(((6aS,8S)-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro -6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)3-methyl-2-oxa-8-azaspiro[ 4,5]Decan-4-yl)carbamate (35 mg, 0.053 mmol, 1.00 equiv) in DCM (1.80 ml) was added dropwise with TFA (0.15 ml) at room temperature. The resulting mixture was stirred for 2 hours at room temperature and then made alkaline to pH=8 aq. ammonium hydroxide solution. The organic solvent was removed in vacuo and the residue was purified by pre-HPLC to give the title compound (7 mg, 23.6%) as a white solid. MS (ER, m/z): [M+1]+=559.3.

Example 32.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)-8- ((2-methoxyethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio) pyrazin-2yl)methanol.

The title compound was synthesized by workup similar to that described in Example 31, steps 6-8 using 1-bromo-2-methoxyethane in step 6. MS (ES, m/z): [M+1] ⁺ =573, 3.

Example 33.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)-8- ((cyclopropylmethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin- 2yl)methanol.

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The title compound was synthesized by workup similar to that described in Example 31, steps 6-8 using (bromomethyl)cyclopropane in step 6. MS (ES, m/z): [M+1]+=569.3.

Example 34.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)-8- ((oxetan-3ylmethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio) pyrazin-2yl)methanol.

The title compound was synthesized by workup similar to that described in Example 31, steps 6-8 using 3-(bromomethyl)oxetane in step 6. MS (ES, m/z): [M+1]+=585.3 .

Example 35.

Synthesis of formic acid; (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((S)2-amino-6a, 7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2yl)methanol.

Step 1: 2-fluoro-4-iodo-6-nitropyridin-3-ol.

To a stirred solution of 2-fluoro-4-iodopyridin-3-ol (100 mg, 0.418 mmol, 1.00 equiv) and Bu ₄ NNO ₃ (191 mg, 0.628 mmol, 1.50 equiv) in DCM (2.0 ml ) add TPCA (0.10 ml, 0.595 mmol, 1.42 equiv) dropwise at 0°C under nitrogen, and the resulting mixture is stirred for 1 hour at 0°C. The reaction is quenched with saturated NH ₄ Cl (aq) at 0° C. and the resulting mixture is extracted with EtOAc. The combined organic layers are washed with water and brine and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE/EtOAc (5:1) to give the title compound (10 mg, 8.4%) as a yellow solid.

Step 2: tert-butyl (S)-2-(((2-fluoro-4-iodo-6-nitropyridin-3-yl)oxy)methyl)pyrrolidine-1carboxylate.

To a stirred solution of 2-fluoro-4-iodo-6-nitropyridin-3-ol (170 mg, 0.599 mmol, 1.00 equiv), tert-butyl(2S)-2-(hydroxymethyl)pyrrolidin-1-carboxylate (180 mg, 0.898 mmol, 1.50 equiv) and

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PPh ₃ (235 mg, 0.9 mmol, 1.5 equiv) in THF (2 ml) add DIAD (181 mg, 0.9 mmol, 1.5 equiv) dropwise at 0°C under nitrogen atmosphere. The resulting mixture was stirred overnight at 60°C under a nitrogen atmosphere and then quenched with water at room temperature. The resulting mixture was extracted with EtOAc and the combined organic layers were washed with water and brine and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-30%) to give the title compound (50 mg, 17.9%) as a yellow oil.

Step 3: (S)-2-fluoro-4-iodo-6-nitro-3-(pyrrolidin-2-ylmethoxy)pyridine hydrochloride.

Add tert-butyl (S)-2-(((2-fluoro-4-iodo-6nitropyridin-3-yl)oxy)methyl)pyrrolidine-1-carboxylate (50 mg, 0.107 mmol, 1) to an 8 ml hermetically sealed tube. .00 equiv) and a solution of 4 M HCl in 1,4-dioxane (0.50 ml, 2.0 mmol, 18.69 equiv) at 0°C. The reaction solution was stirred for 1 hour at room temperature and then concentrated under reduced pressure. The residue was triturated with Et ₂ O to obtain the crude title product (30 mg) as a yellow oil, which was used in the next step without further purification.

Step 4: (S)-4-iodo-2-nitro-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazine.

NO ₂

Add (S)-2-fluoro-4-iodo-6-nitro-3-(pyrrolidin-2ylmethoxy)pyridine hydrochloride (30 mg, crude), K ₂ CO ₃ (34 mg, 0.246 mmol) to an 8 ml hermetically sealed tube. and EtOH (0.30 ml) at room temperature. The resulting mixture was stirred for 2 hours at 60°C and then cooled to room temperature.

The resulting mixture was extracted with EtOAc and the combined organic layers were washed with water and brine and dried over anhydrous Na ₂ SO ₄ . After filtration, the filtrate is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE/EtOAc (5:1) to give the title compound (13 mg, 2 steps, 34.6%) as a yellow solid.

Step 5: (S)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]nirrolo[1,2-d][1,4]oxazin-2-amine.

Mixture of (S)-4-iodo-2-nitro-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazine (13 mg , 0.037 mmol, 1.0 equiv), NH ₄ Cl (4 mg, 0.075 mmol, 2.0 equiv) and Fe (4 mg, 0.075 mmol, 2.0 equiv) in EtOH (0.10 ml) and H ₂ O (0.10 ml) is stirred for 2 hours at 60°C. The mixture is cooled to room temperature and filtered. The filter cake is washed with EtOAc and the filtrate is washed with water and brine, dried over anhydrous Na ₂ SO ₄ After filtration, the filtrate is concentrated under reduced pressure and the residue is purified by silica gel column chromatography, eluted with EtOAc/PE (0-100%) to obtain the title compound (9 mg, 75.8%) as a yellow solid.

Stage 6: formic acid; (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6(((S)-2-amino-6a, 7,8,9-tetrahydro-6H-πpyrido[3,2-b]πpyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2yl)methanol.

The title compound was synthesized by workup similar to that described in Example 5, Steps 7-8 using (S)-4-iodo-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]πpyrrolo [1,2d][1,4]oxazin-2-amine at step 7. MS (ER, m/z): [M+1]+=500.3.

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Example 36 and 37.

Synthesis of (3-((S)-5-amino-5,7-dihydrospiro[cyclopenta[b]niridin-6,4'-niperidin]-1'-yl)-6-(((6aS,8S)-8 (methoxymethyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2yl)methanol [36] and (3-((S)-5-amino-5,7-dihydrospiro[cyclopenta[b]pyridine-6,4'-piperidin]-1'-yl)-6(((6aS,8R) -8-(methoxymethyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4yl)thio)pyrazin-2- yl)methanol [37].

37

Step 1: 2-ethylhexyl 3-(((6aS)-8-(methoxymethyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2d] [1,4 ]oxazin-4-yl)thio)propanoate.

To a stirred mixture of (6aS)-4-iodo-8-(methoxymethyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2b]pyrrolo[1,2-d][1,4]oxazine ( 500 mg, 1.444 mmol, 1.00 equiv), 2-ethylhexyl 3-sulfanylpropanoate (378 mg, 1.733 mmol, 1.20 equiv), Pd ₂ (dba) ₃ (130 mg, 0.142 mmol, 0.10 equiv) and Xanthos (85 mg, 0.147 mmol, 0.10 equiv) in 1,4-dioxane (7.5 ml) was added to DIEA (560 mg, 4.333 mmol, 3.00 equiv) at room temperature under nitrogen and the resulting mixture was stirred. for 2 hours at 85°C. After cooling to rt, the reaction is quenched with water and extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated and the residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-60%) to give the title compound (500 mg, 79.3% ) as a light yellow oil.

Step 2: (6aS)-8-(methoxymethyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazine4-thiolate sodium .

To a stirred solution of 2-ethylhexyl 3-(((6aS)-8-(methoxymethyl)-6a,7,8,9-tetrahydro-6Hpyrido[3,2-b]pyrrolo[1,2-d][1,4 ]oxazin-4-yl)thio)propanoate (500 mg, 1.145 mmol, 1.00 equiv) in MeOH (10.0 ml) add NaOMe (68 mg, 1.260 mmol, 1.10 equiv) at room temperature. After stirring for 5 hours at room temperature, the reaction mixture was concentrated in vacuo and the residue triturated with Et ₂ O to obtain the title compound (220 mg, 70%) as a yellow solid.

Step 3: (3-chloro-6-(((6aS)-8-(methoxymethyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2d] [ 1 ,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol.

To a stirred mixture of (6aS)-8-(methoxymethyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2b]pyrrolo[1,2-d][1,4]oxazine-4-thiolate sodium (100 mg, 0.365 mmol, 1.00 equiv), (6-bromo-3-chloropyrazin2-yl)methanol (90 mg, 0.40 mmol, 1.10 equiv), Pd ₂ (dba) ₃ (33 mg, 0.036 mmol, 0.10 equiv) and Xantphos (21 mg, 0.036 mmol, 0.10 equiv) in 1,4-dioxane (2 ml) add DIEA (141 mg, 1.09 mmol, 3.0 equiv) at room temperature in a nitrogen atmosphere. After stirring for 2 hours at 85°C, the reaction mixture was concentrated in vacuo and the residue was purified by silica gel column chromatography, eluting with EtOAc/PE (0-60%), to give the title compound (105 mg, 72.9% ) in the form of a light yellow oil.

Step 4: (3-((S)-5-amino-5,7-dihydrospiro[cyclopenta[b]pyridine-6,4'-piperidin]-1'-yl)-6(((6aS,8S)- 8-(methoxymethyl)-6a,7,8,9-tetrahydro-6H-nirido[3,2-b]pyrrolo[1,2-d][1,4]oxαzin-4yl)thio)pyrazin-2-yl )methanol and (3-((S)-5-amino-5,7-dihydrospiro[cyclopenta[b]pyridine-6,4'-piperidin]-1'yl)-6-(((6aS,8R)- 8-(methoxymethyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4yl)thio)pyrazin-2-yl )methanol.

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To the stirred mixture (3-chloro-6-(((6aS)-8-(methoxymethyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2b]pyrrolo[1,2-d][1 ,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol (50 mg, 0.127 mmol, 1.00 equiv) and (S)-5,7dihydrospiro[cyclopenta[b]pyridine-6,4'- piperidine]-5-amine trihydrochloride (50 mg, 0.159 mmol, 1.25 equiv) in ACN (2.0 ml) add DIEA (82 mg, 0.63 mmol, 5.0 equiv) at room temperature and the resulting mixture is stirred at 80°C for 48 hours. After cooling to room temperature, the reaction mixture is filtered and the filter cake is washed with MeOH and the combined filtrate is concentrated and the residue is purified by pre-HPLC to give the title compound. MS (ER, m/z): [M+1]+=562.3.

Example 38.

Synthesis of (3-((S)-5-amino-13-oxa-9-azadispiro[3,1,56,24]tridecan-9-yl)-6-(((6aS,8S)-8-(methoxymethyl )6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol.

Solution of (3-chloro-6-(((6aS,8S)-8-(methoxymethyl)-6α,7,8,9-tetraαhydro-6H-nirido[3,2-b]pyrrolo[1,2d][1 ,4]oxazin-4-yl)thio)nirasin-2-yl)methanol (35 mg, 0.089 mmol, 1.00 equiv), (S)-13-oxa-9azadispiro[3,1,5 ⁶ ,2 ⁴ ]tridecan-5-amine dihydrochloride (29 mg, 0.106 mmol, 1.20 equiv) and DIEA (57 mg, 0.443 mmol, 5.00 equiv) in CH ₃ CN (0.56 ml) were stirred for 16 hours at 80 °C. The reaction mixture was purified by pre-HPLC to give the title compound (9.8 mg, 18.4%) as a yellow solid. MS (ER, m/z): [M+1]+=555.2.

Example 39 and 40.

Synthesis of (3-((S)-5-amino-13-oxa-9-azadisniro[3,1,56,24]tridecan-9-yl)-6-(((6aS,8S)-8-(( methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)nirazin-2yl )methanol [39].

The title compound was synthesized by treatment analogous to that described in Examples 36 and 37 from (6aS,8S)-4-iodo-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido [3,2b]n-pyrrolo[1,2-d][1,4]oxazine and (S)-13-oxa-9-azadispiro[3,1,5 ⁶ ,2 ⁴ ]tridecan-5-amine dihydrochloride. MS (ER, m/z): [M+1]+=585.3.

(3-((3S,4S)-4-Amino-3-methyl-2-oxa-8-azacniro[4,5]decan-8-yl)-6-(((6aS,8S)-8-( ((methoxymethoxy)methoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio) pyrazin-2-yl)methanol [40]

isolated as a by-product.

Example 41.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)-8( hydroxymethyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)πyrazin-2yl)methanol.

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Step 1: (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)- 8-(hydroxymethyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2 -yl)methanol.

During the synthesis of Example 31, compound 42 was also isolated as a by-product. MS (ER, m/z): [M+1]+=515.3.

Example 42.

Synthesis of (3S,4S)-8-(5-(((6aS,8S)-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2b]nirrolo[1 ,2-d][1,4]oxazin-4-yl)thio)nirazin-2-yl)-3-methyl-2-oxa-8-azacyro[4,5]decane-4-amine.

Step 1: (6aS,8S)-4-((5-chloropyrazin-2-yl)thio)-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6Hpyrido[3,2-b ]pyrrolo[1,2-d][1,4]oxazine.

Mixture of (6aS,8S)-4-iodo-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-πpyrigo[3,2-b]πpyrrolo[1,2d][1,4 ]oxazine (90 mg, 0.239 mmol, 1.00 equiv), sodium 5-chloropyrazine-2-thiolate (61 mg, 0.36 mmol, 1.51 equiv) (see WO 2016/203406), Pd ₂ (dba ) ₃ (22 mg, 0.024 mmol, 0.10 equiv), Xantphos (14 mg, 0.024 mmol,

0.10 equiv) and DIEA (93 mg, 0.72 mmol, 3.0 equiv) in THF (2 ml) were stirred for 0.5 h at 60°C under nitrogen atmosphere. The residue was purified by silica gel column chromatography to give the title compound (93 mg, 98.5%) as a brown oil. MS (ER, m/z): [M+1] ⁺ =395.2.

Step 2: (3S,4S)-8-(5-(((6aS,8S)-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2b]pyrrolo [1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4,5]decane-4-amine.

Solution of (6aS,8S)-4-((5-chloropyrazin-2-yl)thio)-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6Hpyrido[3,2-b]pyrrolo [1,2-d][1,4]oxazine (40 mg, 0.101 mmol, 1.00 equiv), (3S,4S)-3-methyl-2-oxa-8azaspiro[4,5]decane-4- amine dihydrochloride (42 mg, 0.173 mmol, 1.70 equiv) and DIEA (65 mg, 0.50 mmol,

5.00 equiv) in CH3CN (0.40 ml) was stirred for 16 hours at 100°C. The residue was purified by pre-HPLC to give the title compound (16 mg, 29.9%) as an off-white solid. MS (ER, m/z): [M+1] ⁺ =529.3.

Example 43.

Synthesis of (3S,4S)-8-(5-(((6aS,8S)-8-(methoxymethyl)-6a,7,8,9-tetrahydro-6H-pyrudo[3,2-b]pyrrolo[1, 2d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4,5]decane-4-amine.

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The title compound was synthesized by treatment similar to that described in Example 42. MS (ES, m/z): [M+1]+=499.3.

Example 44.

Synthesis of (3S,4S)-8-(6-amino-5-(((6aS,8S)-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2b ]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4,5]decane-4-amine .

Step 1: 6-chloro-3-(((6aS,8S)-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2b]pyrrolo[1,2- d][1,4]oxazin-4-yl)thio)pyrazin-2-amine.

Mixture of (6aS,8S)-4-iodo-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2d][1,4 ]oxazine (90.00 mg, 0.239 mmol, 1.00 equiv), sodium 3-amino-5-chloropyrazine-2-thiolate (60 mg, 0.359 mmol, 1.5 equiv) (see WO 2015/107494), Pd ₂ (dba) ₃ (22 mg, 0.024 mmol, 0.1 equiv), Xantphos (14 mg, 0.024 mmol, 0.1 equiv) and DIEA (93 mg, 0.72 mmol, 3.0 equiv) in THF (2.00 ml) was stirred for 0.5 h at 60°C under nitrogen atmosphere. The residue was purified by silica gel column chromatography to give the title compound (93 mg, 98.9%) as a brown oil.

Step 2: (3S,4S)-8-(6-amino-5-(((6aS,8S)-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6Hpyrido[3,2 -b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4,5]decane-4amine .

Solution of 6-chloro-3-(((6aS,8S)-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2b]pyrrolo[1,2-d] [1,4]oxazin-4-yl)thio)pyrazin-2-amine (40 mg, 0.1 mmol, 1.00 equiv), (3S,4S)-3-methyl-2oxa-8-azaspiro[4 ,5]decane-4-amine dihydrochloride (42 mg, 0.17 mmol, 1.76 equiv) and DIEA (65 mg, 0.5 mmol, 5.1 equiv) in CH3CN (0.40 ml) stir for 16 hours at 100°C. The residue was purified by pre-HPLC to give the title compound (16 mg, 30%) as an off-white solid. MS (ER, m/z): [M+1]+=544.3.

Example 45.

Synthesis of (3S,4S)-8-(6-amino-5-(((6aS,8S)-8-(methoxymethyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2b]pyrrolo[ 1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4,5]decane-4-amine.

The title compound is synthesized by processing similar to that described in

- 66 044940 example 44. MS (ER, m/z): [M+1]+=514.3.

Example 46.

Synthesis of (S)-1'-(5-(((6aS,8S)-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2b]pyrrolo[1, 2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)-5,7-dihydrospiro[cyclopenta[b]pyridin-6,4'piperidine]-5-amine.

Solution of (6aS,8S)-4-((5-chloropyrazin-2-yl)thio)-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6Hpyrido[3,2-b]pyrrolo [1,2-d][1,4]oxazine (40 mg, 0.10 mmol, 1.0 equiv), (S)-5,7dihydrospiro[cyclopenta[b]pyridine-6,4'-piperidine]- 5-amine trihydrochloride (48 mg, 0.15 mmol, 1.5 equiv) and DIEA (65 mg, 0.50 mmol, 5.00 equiv) in CH ₃ CN (0.40 ml) were stirred for 16 h at 100°C. The reaction solution was purified by pre-HPLC to give the title compound (12.0 mg, 21.2%) as an off-white solid, MS (ES, m/z): [M+1]+=562.3.

Example 47.

Synthesis of (S)-1'-(6-amino-5-(((6aS,8S)-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2b] pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)-5,7-dihydrospiro[cyclopenta[b]pyridine-6,4'piperidine]-5-amine .

Solution of 6-chloro-3-(((6aS,8S)-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2b]pyrrolo[1,2-d] [1,4]oxazin-4-yl)thio)pyrazin-2-amine (40 mg, 0.098 mmol, 1.00 equiv), (S)-5,7dihydrospiro[cyclopenta[b]pyridine-6,4'- piperidine]-5-amine trihydrochloride (46 mg, 0.147 mmol, 1.50 equiv) and DIEA (63 mg, 0.487 mmol, 5.0 equiv) in CH ₃ CN (0.40 ml) stir for 16 hours at 100 °C. The reaction solution was purified by pre-HPLC to give the title compound (16.0 mg, 28.4%) as an off-white solid. MS (ER, m/z): [M+1]+=577.3.

Example 48.

Synthesis of (3-((S)-5-amino-5,7-dihydrospiro[cyclopenta[b]pyridine-6,4'-piperidine]-1'-ul)-6-(((6aS,8S)-8 ((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo-[1,2-d][1,4]oxazin-4yl)thio)pyrazine-2 -yl)methanol.

To a stirred solution of (S)-5,7-dihydrospiro[cyclopenta[b]pyridine-6,4'-piperidine]-5amine trihydrochloride (28 mg, 0.09 mmol, 1.53 equiv) and DIEA (38 mg, 0.294 mmol, 5.0 equiv) to ACN (0.5 ml) add (3-chloro-6-(((6aS,8S)-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro- 6H-pyrido[3,2b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol (25 mg, 0.059 mmol, 1.00 equiv) at room temperature in a nitrogen atmosphere. The resulting mixture was stirred overnight at 100°C, and the mixture was cooled to room temperature. The crude product was purified by pre-HPLC to give the title compound (6.5 mg, 18.67%) as a white solid. MS (ER, m/z): [M+1]+=592.3.

Example 49.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8S)-8- ((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)-5 -methylpyrazin2-yl)methanol.

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The title compound was synthesized by treatment similar to that described in Example 31. MS (ES, m/z): [M+1] ⁺ =573.3.

Example 50.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8R)-8- ((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin- 2yl)methanol.

Step 1: (3-chloro-6-(((6aS)-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido-[3,2b]pyrrolo[1,2- d] [1,4]oxazin-4-yl)thio)pyrazin-2-yl)methanol.

Into the mixture (6aS)-4-iodo-8-((methokCymetokCu)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2d][1,4] oxazine (1.00 g, 2.658 mmol, 1.00 equiv) and sodium 5-chloro-6-(hydroxymethyl)pyrazine-2-thiolate (0.78 g, 3.93 mmol, 1.48 equiv) in THF ( 10.00 ml) add Xantphos (0.15 g, 0.26 mmol, 0.10 equiv) and DIEA (1.02 g, 7.892 mmol, 2.97 equiv) at room temperature. Pd ₂ (dba) ₃ (0.12 g, 0.13 mmol, 0.05 equiv) was then added to the solution under nitrogen and the resulting mixture was stirred for 1 hour at 60° C. under nitrogen. The resulting mixture was cooled to rt and filtered, and the filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography, eluted with PE/EA (0~100%) to give the product (790 mg, 69.9%).

Step 2: (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aS,8R)- 8((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4yl)thio)pyrazin-2 -yl)methanol.

To a mixture of (3S,4S)-3-methyl-2-oxa-8-azaspiro[4,5]decane-4-amine dihydrochloride (730 mg, 3.002 mmol, 1.72 equiv) and DIEA (1.30 g, 10.06 mmol, 5.77 equiv) to ACN (5 ml) add (3-chloro-6-(((6aS)-8((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H- pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4yl)thio)pyrazin-2-yl)methanol (740 mg, 1.742 mmol, 1.00 equiv) at room temperature. The resulting mixture was stirred for 16 hours at 60°C under nitrogen and then purified by pre-HPLC and chiral HPLC to obtain the product. MS (ER, m/z): [M+1]+=559.3.

Example 51 and 52.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aR,8S)-8- ((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio)pyrazin- 2yl)methanol (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-(((6aR,8R)- 8 ((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4yl)thio)pyrazin-2 -yl)methanol.

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Step 1: (6aR)-4-iodo-8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,20][1,4 ]oxazine.

The title compound is synthesized by treatment similar to that described in Examples 29-30, Steps 1-6.

Step 2: (3-((3S,4S)-4-amino-3-methyl-2-oxα-8-azaspiro[4,5]decαn-8-yl)-6-(((6aR,8S)- 8-((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)thio) pyrazin-2yl)methanol and (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro-[4,5]decan-8-yl)-6-(((6aR ,8R)-8((methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4yl)thio )pyrazin-2-yl)methanol.

A mixture of the title compounds is synthesized by processing similar to that described in example 31, steps 5-8. The product was then purified by HPLC to give compounds 51 and 52. Compound 51: MS (ES, m/z): [M+1]+=559.3. Compound 52: MS (ER, m/z): [M+1] ⁺ =559.3.

Example 53.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azacniro[4,5]decan-8-yl)-6-((6aS,8S)-8-( methoxymethyl)6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)pyrazin-2-yl)methanol.

Step 1: ((6aS,8S)-8-(methoxymethyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo|1,2-d]-[1,4] oxazin4-yl)boronic acid.

To a stirred solution of (6aS,8S)-4-iodo-8-(methoxymethyl)-6α,7,8,9-tetrahydro-6H-pyrido[3,2b]pyrrolo[1,2-d][1,4] oxazine (100 mg, 0.289 mmol, 1.00 equiv), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2dioxaborolane) ( 110 mg, 0.433 mmol, 1.5 equiv), AcOK (57 mg, 0.581 mmol, 2.0 equiv) in toluene (2 ml) add Pd(dppf)Cl ₂ (12 mg, 0.0164 mmol, 0.057 equiv) at room temperature in an N2 atmosphere. The resulting mixture was stirred for 16 hours at 100°C under N2 atmosphere, and the resulting mixture was used immediately in the next step without further purification.

Step 2: tert-butyl ((3S,4S)-8-(3-(hydroxymethyl)-5-((6aS,8S)-8-(methoxymethyl)-6a,7,8,9tetrahydro-6H-nirido[3 ,2-b]nirrolo[1,2-d][1,4]oxazin-4-yl)pyrazin-2-yl)-3-methyl-2-oxa-8azaspiro[4,5]decan-4-yl )carbamate.

Vos

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To the mixture ((6aS,8S)-8-(methoxymethyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d] [1,4]oxazin- 4yl)boronic acid in toluene from the last stage add K ₂ CO ₃ (120 mg, 0.87 mmol, 3.00 equiv), tert-butyl ((3S,4S)-8-(5-bromo-3-(hydroxymethyl )pyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4,5]decan-4yl)carbamate (132 mg, 0.290 mmol, 1.00 equiv), Pd(dppf)Cl ₂ (12 .00 mg, 0.064 mmol, 0.057 equiv) and H ₂ O (0.2 ml) under N ₂ . After stirring for 3 hours at 100°C, the resulting mixture is concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with EA to give the title compound (70 mg, 40.7%, 2 steps) as a light yellow oil. MS (ER, m/z): [M+1]=597.3.

Step 3: (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-((6aS,8S)-8 (methoxymethyl)-6a,7,8,9-tetrahydro-6H-nirido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)pyrazin-2yl)methanol.

A mixture of tert-butyl ((3S,4S)-8-(3-(hydroxymethyl)-5-((6aS,8S)-8-(methoxymethyl)-6a,7,8,9-tetrahydro6H-pyrido[3,2 -b]pyrrolo[1,2-d][1,4]oxazin-4-yl)pyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4,5]decane-4yl)carbamate (70 mg, 0.117 mmol, 1.00 equiv) in DCM (3 mL) and TFA (1 mL) stirred at room temperature for 3 hours. The resulting mixture was concentrated and purified by pre-HPLC to give the title compound (17 mg , 29.3%). MS (ER, m/z): [M+H]+=497.3.

Example 54.

Synthesis of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4,5]decan-8-yl)-6-((6aS,8S)-8-( (methoxymethoxy)methyl)-6a,7,8,9-tetrahydro-6H-pyrido[3,2-b]pyrrolo[1,2-d][1,4]oxazin-4-yl)pyrazin-2yl)methanol .

The title compound was synthesized by treatment similar to that described in Example 54. MS (ES, m/z): [M+H]+=527.3.

Biological analyses.

SHP2 allosteric inhibition assay.

SHP2 possesses two N-terminal Src homology 2 (SH2) domains, a central protein tyrosine phosphatase (PTP) domain and a C-terminal tail. In the basal state, SHP2 is self-inhibited and the access of substrates to the catalytic site is blocked by intermolecular interactions between the SH2 domains and the PTP domain. When bis-tyrosiphosphorylated peptides bind to the SH2 domain of SHP2, the PTP domain becomes available for substrate recognition and reaction catalysis, and SHP2 is allosterically activated. The catalytic activity of SHP2 can be measured using the fluorogenic artificial substrate DiFMUP.

Phosphatase reactions are performed at room temperature in 384-well black polystyrene plates (Greiner Bio-One, Cat #784076) using assay buffers containing 60 mM HEPES, pH 7.2, 75 mM NaCl, 75 mM KCl, 1 mM EDTA, 0.05% P-20 and 5 mM DTT.

0.33 nM SHP2 was co-incubated with 0.5 µM biphos-IRS1 peptide (sequence: H2NLN(pY)IDLDLV(dPEG8)LST(pY)ASINFQK-amide) and different concentrations of compounds for 3060 min at room temperature. The reaction is then initiated by adding DiFMUP surrogate substrate (Invitrogen, Cat# D6567, 100 µM final volume).

Real-time conversion of DiFMUP to DiFMU (6,8-difluoro-7-hydroxyl-4methylcoumarin) is measured every 5 min for 30 min using a microplate reader (CLARIOstar, BMG Labtech) with excitation and emission wavelengths of 340 nm and 450 nm , respectively. Baseline response rates are determined by linear fitting of the data, and inhibitor dose response curves are analyzed using normalized IC ₅₀ regression curve fitting with normalization to control.

The IC ₅₀ value for the compounds numbered in the table. 1 with connections are presented below in table. 4, where IC ₅₀ were measured as 100 nM or lower.

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Table 4

Compound No. from Table 1 Structure Shp2 1C ₅₀ (nM) 1 Wed & 2 K> 3 2 Τη n 7 ΝΗ _Ζ no ..... 4 3 ^Ρ ^γ\ ³ Ύ^“ N^J Ν lk _N ^ but/ h ₂ n \ 13 4 f/L HO N%o h ₂ n \ 5 5 II ΊΓ ⁿ N^ _n -X but/ %j2° h ₂ n \ 8 6 ^Me0 CCVs^ ΙΙΊ Γ^ ^Ν but/ h ₂ n \ 6 7 ^ho <Cv^ _n but/ h ₂ n \ 6

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8 ^Ν >U'n-b>b ^o N ₂ H\ 8 9 o-Yl ? ^N O ΙΑλ nh ₂ Y Y>..... 8 10 ^N Y nAzk NH ₂ J 4ύ..... 10 AND >>-αχ ₅ _ _Ν 1—' Η 11· ^Ν 4/ ΝΗ ₂ γ>..... 14 12 V ^Ν Υ _ΝΗ ₂ ^X ohQ)..... 12 13 Γ\ Τ Ί Ίτ ^ν ο- ³ νη ₂ ho ³ 4Q..... 8 14 rCw, \ Ί Ν °- ϋ hcJ <Υ° η ₂ ν \ 5

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15 .. / ^N xi> NH ₂ Meo γ n η _? ww..... 10 16 a. /\Z \/(N 4 CD € 3 17 ΥΎ r J NSG ^O° h ₂ n \ 2 18 19 4/3^ ⁰ HO*\-N. II ^N nh ₂ ho^ ^1χ Ηθ*..... *4/3^° ΚοΆ^Ν , z<x _z s, _z .-s Η ίΓ ^N NH ₂ A ..... one of 21 and 22 has 13 and the other has 8 20 “-ciSLs^ ji n “Al ho^ ^lx ^t/° H ₂ hr\ AND 21 0=s-< I 1 ^ιΎ ΥυΥ ^N > J 43 n ₂ gG\ 13

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22 I z 0^0 £ 45 4? 6 23 ° nh ₂ J 45..... 8 24 J ' 0 ¹ / ² \ 4 > / \ ^z \ / 10 25 Γ^Νι-ΥΥ^? 4' 0 ^Ν γ4γ^ _Ν Ύ Y h ₂ n \ eleven 26 and 27 Q 0 rt-o 4 co " o4 : "Ξ one of 26 and 27 has 10 and the other has 8 28 0 ^v N N^ ^{1 N} aA _n ^\ NH ₂ J 43..... ^-o 13 29 £ £'i 7 Φ 3.6

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thirty /Uo ' \^N Ail MeO ]|Υ mU-^l νη ₂ no ..... U 3.8 31 o^^ ^N yV ^s yy ⁴ h> η ₂ ν \ 4.2 32 уУ^о °\ ⁿ yV^ ⁿ / Μ ^N y^ _N y OMe ) L У>„ ^but %L· ⁰ Η,Ν \ 4 33 Zy^o 7 ·<%Χ n ₂ /\ 2.3 34 yу^o °C^ ⁿ y^ ^s ^ ⁿ Usk n ₂ g/\ 6.2 35 lu^o Y Y N ncoon ^N y^N^ Y2 NH ₂ |_| _P / ..... pi ( / ^-o 18

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36 and 37 ¹ l 3 Wed 4° s ,γ Mi 'Μ ? ζ sD ^m One of 36 and 37 has 2.7 and the other has 4.2 38 /Y^o\hY yy NH ₂ J 4χ> 5.7 39 ζΎ^ο γν ^Αγ-s mJ E IJ/ry tM ho ^; 4γ 7.0 40 Ζ-γ^ο °\ ⁿ W ^s y^n 2 N >k _N ^ MeO^ ^H0 ^ \Q h ₂ n \ 5.9 41 /Y^O H0^Y ^N yV ^s y^ _N nZ «j y> H ₂ N ^r \ 5.0 42 SL fl t^ / \ 5.9

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p-ERK cell assay.

Detroit562 cells are seeded in a 96-well plate and cultured overnight (30,000 cells per well, 200 μl total volume). In the morning, cells are treated with compounds as described at an initial concentration of 10 μM and 1/2 log dilution to 1 nM for 2 hours at 37°C. DMSO treatment serves as a control. p-ERK is then measured using the AlphaLISA® SureFire® Ultra™ p-ERK 1/2 (Thr202/Tyr204) Assay Kit (PerkinElmer, ALSU-PERK-A500) following the instructions. Briefly, the medium is removed and 50 μl of IX lysis buffer is added, then incubated on a plate shaker at room temperature for 10 minutes. Then, 10 μl of the lysate was transferred to a white 384-well plate and 5 μl of the acceptor mixture and 5 μl of the donor mixture (both prepared according to the manufacturer's instructions) were added. The plates are wrapped in foil, shaken for 1-2 minutes on a plate reader, and incubated for >2 hours. The signal is then measured on a CLARIOstar® tablet reader. The proportion of inhibition was calculated by taking DMSO treatment as 100% signal, and the IC50 was calculated using Graphpad Prism 7.

Examples of compositions.

The following are typical pharmaceutical formulations containing a compound of formula (PC).

Tablet composition.

The following ingredients are thoroughly mixed and compressed into single scored tablets.

Ingredient Amount per tablet (mg)

Connection as described

Corn starch

Croscarmellose sodium

Lactose

400

120

Magnesium stearate 5

Encapsulated composition.

The following ingredients are thoroughly mixed and loaded into a hard gelatin capsule.

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Ingredient Amount per capsule (mg)

Connection as described200

Dry lactose spray148

Magnesium stearate2

Composition for injection.

Compound as described (eg, Compound 1) in 2% HPMC, 1% Tween 80 in DI water, pH 2.2 with MSA, as needed, to at least 20 mg/ml.

Composition for inhalation.

To prepare a pharmaceutical composition for inhalation, 20 mg of the compound described herein is mixed with 50 mg of anhydrous citric acid and 100 ml of 0.9% sodium chloride solution. The mixture is administered into an inhalation device, such as a nebulizer, which is suitable for inhalation administration.

Gel composition for topical application.

To prepare a pharmaceutical gel composition for topical use, 100 mg of the compound described herein is mixed with 1.75 g of hydroxypropyl cellulose, 10 ml of propylene glycol, 10 ml of isopropyl myristate and 100 ml of purified alcohol USP. The resulting gel mixture is then placed into containers, such as tubes, that are suitable for topical use.

Composition of ophthalmic solution.

To prepare a pharmaceutical ophthalmic solution composition, 100 mg of a compound described herein is mixed with 0.9 g of NaCl in 100 ml of purified water and filtered using a 0.2 micron filter. The resulting isotonic solution is then administered into ophthalmic delivery units, such as eye drop containers, which are suitable for ophthalmic administration.

Nasal spray solution.

To prepare a pharmaceutical solution for nasal spray, 10 g of the compound described herein is mixed with 30 ml of 0.05 M phosphate buffer solution (pH 4.4). The solution is placed in a nasal dispenser designed to dispense 100 µl of spray with each application.

Claims (61)

1. Compound of formula (IIB): R ⁹ R ¹ 5=N /—\ R ^{11 R} Vk /rIZ r3 —Г )=/ νΗ ⁴ —' ^к N—ζ i R ¹⁰ J-I N-I ⁴ R ⁶ Z _R 4 R ⁵ (IIB) or a pharmaceutically acceptable salt thereof, where: where Z is -Y-M; Y is a bond; M is alkyl substituted with -O(alkylene)OR ^a where R ^a is alkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl or heterocyclylalkyl; or Z is methoxymethyloxy, methoxyethyloxy, methoxymethyl, cyclopropylmethyloxymethyl or oxetan-3-ylmethyloxymethyl; or Z is (methoxymethoxy)methoxymethyl; R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen, alkyl or halogen; R ⁵ and R ⁶ are independently selected from hydrogen, alkyl or halogen; L is a bond, O, S, S(O) or S(O) ₂ ; R ⁹ is hydrogen, alkyl, halogen or amino; R ¹⁰ is hydrogen, alkyl, halogen, hydroxy, hydroxyalkyl or -CD2OH; And R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from - 79 044940 where alkyl means a linear saturated monovalent hydrocarbon radical containing from one to six carbon atoms, or a branched saturated monovalent hydrocarbon radical containing from three to six carbon atoms; alkylene means a linear saturated divalent hydrocarbon radical containing from one to six carbon atoms, or a branched saturated divalent hydrocarbon radical containing from three to six carbon atoms; hydroxyalkyl means a linear monovalent hydrocarbon radical containing from one to six carbon atoms, or a branched monovalent hydrocarbon radical containing from three to six carbon atoms, substituted by one or two hydroxy groups, provided that if two hydroxy groups are present, they are not on the same and the same carbon atom; cycloalkyl means a monocyclic saturated monovalent hydrocarbon radical containing from three to ten carbon atoms; heterocyclyl means a saturated monovalent monocyclic or bicyclic ring containing 4-10 ring atoms, in which one, two or three ring atoms are heteroatoms selected from N, O and S(O) _n , where n is an integer from 0 to 2, the remaining ring atoms are C; cycloalkylalkyl means the radical -(alkylene)-R, where R represents cycloalkyl as defined above; heterocyclylalkyl means the radical -(alkylene)-R, where R represents heterocyclyl as defined above. 2. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyloxy, methoxyethyloxy, methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, cyclopropylmethyloxymethyl or oxetan-3-ylmethyloxymethyl. 3. A compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, cyclopropylmethyloxymethyl or oxetan-3-ylmethyloxymethyl. 4. A compound according to claim 3 or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl or cyclopropylmethyloxymethyl. 5. A compound according to claim 4 or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl or methoxyethyloxymethyl. 6. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen and alkyl. 7. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein one of ^R1 , ^R2 , ^R3 and ^R4 is hydrogen, and the remaining three of ^R1 , ^R2 , ^R3 and ^R4 are independently selected from hydrogen, methyl and fluorine. 8. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein three of ^R1 , ^R2 , ^R3 and ^R4 are hydrogen, and the remaining one of ^R1 , ^R2 , ^R3 and ^R4 is selected from hydrogen, methyl and fluoride. 9. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein each of R ¹ , R ² , R ³ and R ⁴ is hydrogen. 10. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein L is S, S(O) or S(O) ₂ . 11. A compound according to claim 10 or a pharmaceutically acceptable salt thereof, wherein L is S. 12. The compound according to claim 10 or a pharmaceutically acceptable salt thereof, where L is S(O) ₂ . 13. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ⁹ is hydrogen, amino or alkyl. 14. The compound according to claim 13 or a pharmaceutically acceptable salt thereof, wherein R ⁹ is hydrogen. 15. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ¹⁰ is hydroxyalkyl. - 80 044940 16. The compound according to claim 15 or a pharmaceutically acceptable salt thereof, wherein R ¹⁰ is hydroxymethyl. 17. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, where Z is methoxymethyloxy, methoxyethyloxy, methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, cyclopropylmethyloxymethyl or oxetan-3-ylmethyloxymethyl; R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen, alkyl and halogen; L is S, S(O) or S(O) ₂ ; R ⁹ is hydrogen, amino or alkyl; and R ¹⁰ is hydroxyalkyl. 18. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from 19. A compound according to claim 18 or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon to which they are attached form a ring selected from the with atom 20. A compound according to claim 18 or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon to which they are attached form a ring selected from 21. A compound according to claim 17 or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from 22. A compound according to claim 21 or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from 23. A compound according to claim 21 or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from 24. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyloxy, methoxyethyloxy, methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, cyclopropylmethyloxymethyl or oxetan-3-ylmethyloxymethyl; R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen, alkyl and halogen; L is S, S(O) or S(O) ₂ ; R ⁹ is hydrogen, amino or alkyl; R ¹⁰ is hydroxyalkyl; And R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from - 81 044940 25. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, where Z is methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl or methoxyethyloxymethyl; R ¹ , R ² , R ³ and R ⁴ are hydrogen; L is S, S(O) or S(O) ₂ ; R ⁹ is hydrogen; R ¹⁰ is hydroxyalkyl; And R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from R and 26. A compound according to claim 25 or a pharmaceutically acceptable salt thereof, wherein the R carbon to which they are attached forms a ring selected from together with an atom 27. A compound according to claim 25 or a pharmaceutically acceptable salt thereof, wherein the R ¹¹ carbon to which they are attached forms a ring selected from R ¹² together with an atom 28. The compound according to claim 25 or a pharmaceutically acceptable salt thereof, where L is S. 29. A pharmaceutical composition containing a compound according to claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. 30. Compound of formula (IIB): or a pharmaceutically acceptable salt thereof, wherein Z is -Y-M; Y is a bond; M is alkyl substituted with alkoxy or -O(alkylene)OR ^a where R ^a is alkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl or heterocyclylalkyl; or Z is alkoxy, cycloalkylmethyloxymethyl, heterocyclyloxy, heterocyclylmethyloxymethyl; where if Z is alkoxy, it is optionally substituted with methoxy; R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen, alkyl or halogen; R ⁵ and R ⁶ are independently selected from hydrogen, alkyl or halogen; L is a bond, O, S, S(O) or S(O) ₂ ; R ⁹ is hydrogen, alkyl, halogen or amino; R ¹⁰ is hydrogen, alkyl, halogen, hydroxy, hydroxyalkyl or -CD2OH; And R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from - 82 044940 where alkyl means a linear saturated monovalent hydrocarbon radical containing from one to six carbon atoms, or a branched saturated monovalent hydrocarbon radical containing from three to six carbon atoms; alkylene means a linear saturated divalent hydrocarbon radical containing from one to six carbon atoms, or a branched saturated divalent hydrocarbon radical containing from three to six carbon atoms; hydroxyalkyl means a linear monovalent hydrocarbon radical containing from one to six carbon atoms, or a branched monovalent hydrocarbon radical containing from three to six carbon atoms, substituted by one or two hydroxy groups, provided that if two hydroxy groups are present, they are not on the same and the same carbon atom; cycloalkyl means a monocyclic saturated monovalent hydrocarbon radical containing from three to ten carbon atoms; heterocyclyl means a saturated monovalent monocyclic or bicyclic ring containing 4-10 ring atoms, in which one, two or three ring atoms are heteroatoms selected from N, O and S(O)n, where n is an integer from 0 to 2, the remaining ring atoms are C; cycloalkylalkyl means the radical -(alkylene)-R, where R represents cycloalkyl as defined above; heterocyclylalkyl means the radical -(alkylene)III, where R represents heterocyclyl as defined above. 31. A compound according to claim 30 or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyloxy, methoxyethyloxy, methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, cyclopropylmethyloxymethyl or oxetan-3-ylmethyloxymethyl. 32. A compound according to claim 31 or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, cyclopropylmethyloxymethyl or oxetan-3-ylmethyloxymethyl. 33. A compound according to claim 32 or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl or cyclopropylmethyloxymethyl. 34. A compound according to claim 33 or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl or methoxyethyloxymethyl. 35. A compound according to claim 30 or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen and alkyl. 36. The compound of claim 35 or a pharmaceutically acceptable salt thereof, wherein one of ^R1 , ^R2 , ^R3 and ^R4 is hydrogen and the remaining three of ^R1 , ^R2 , ^R3 and ^R4 are independently selected from hydrogen, methyl and fluorine. 37. A compound according to claim 35 or a pharmaceutically acceptable salt thereof, wherein three of ^R1 , ^R2 , ^R3 and ^R4 are hydrogen, and the remaining one of ^R1 , ^R2 , ^R3 and ^R4 is selected from hydrogen, methyl and fluoride. 38. The compound according to claim 35 or a pharmaceutically acceptable salt thereof, wherein each of R ¹ , R ² , R ³ and R ⁴ is hydrogen. 39. A compound according to claim 30 or a pharmaceutically acceptable salt thereof, wherein L is S, S(O) or S(O) ₂ . 40. The compound according to claim 39 or a pharmaceutically acceptable salt thereof, where L is S. 41. A compound according to claim 39 or a pharmaceutically acceptable salt thereof, wherein L is S(O) ₂ . 42. A compound according to claim 30 or a pharmaceutically acceptable salt thereof, wherein R ⁹ is hydrogen, amino or alkyl. 43. The compound according to claim 42 or a pharmaceutically acceptable salt thereof, wherein R ⁹ is hydrogen. 44. The compound according to claim 30 or a pharmaceutically acceptable salt thereof, wherein R ¹⁰ is hydroxyalkyl. - 83 044940 45. The compound according to claim 44 or a pharmaceutically acceptable salt thereof, wherein R ¹⁰ is hydroxymethyl. 46. The compound according to claim 30 or a pharmaceutically acceptable salt thereof, where Z is methoxymethyloxy, methoxyethyloxy, methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, cyclopropylmethyloxymethyl or oxetan-3-ylmethyloxymethyl; R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen, alkyl or halogen; L is S, S(O) or S(O) ₂ ; R ⁹ is hydrogen, amino or alkyl; And R ¹⁰ is hydroxyalkyl. 47. A compound according to claim 30 or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon to which they are attached form a ring selected from LLAL 48. A compound according to claim 47 or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon to which they are attached form a ring selected from NH ₂ ΝΗ ₂ ΝΗ ₂ 49. The compound according to claim 47 or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon to which they are attached form a ring selected from with atom with atom with atom LLLLL with an atom 50. A compound according to claim 46 or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon to which they are attached form a ring selected from /VW 51. A compound according to claim 50 or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from 52. A compound according to claim 50 or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from 53. A compound according to claim 30 or a pharmaceutically acceptable salt thereof, wherein Z is methoxymethyloxy, methoxyethyloxy, methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl, methoxyethyloxymethyl, cyclopropylmethyloxymethyl or oxetan-3-ylmethyloxymethyl; R ¹ , R ² , R ³ and R ⁴ are independently selected from hydrogen, alkyl or halogen; L is S, S(O) or S(O) ₂ ; R ⁹ is hydrogen, amino or alkyl; R ¹⁰ is hydroxyalkyl; And R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from - 84 044940 54. The compound according to claim 30 or a pharmaceutically acceptable salt thereof, where Z is methoxymethyl, methoxymethyloxymethyl, ethoxymethyloxymethyl or methoxyethyloxymethyl; R ¹ , R ² , R ³ and R ⁴ are hydrogen; L is S, S(O) or S(O) ₂ ; R ⁹ is hydrogen; R ¹⁰ is hydroxyalkyl; And R ¹¹ and R ¹² together with the carbon atom to which they are attached form a ring selected from 55. The compound according to claim 54 or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² of the carbon to which they are attached form a ring selected from together with an atom 56. The compound according to claim 54 or a pharmaceutically acceptable salt thereof, wherein R ¹¹ and R ¹² of the carbon to which they are attached form a ring selected from together with an atom 57. The compound according to claim 54 or a pharmaceutically acceptable salt thereof, where L is S. 58. A pharmaceutical composition containing a compound according to claim 30 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. 59. A compound selected from the group consisting of: - 85 044940 but or a pharmaceutically acceptable salt thereof. 60. The connection according to claim 59, where the connection is - 86 044940 61. Connection by 62. Connection by 63. Connection by 64. Connection by 65. Connection by 66. Connection by /^γ^ο I \ II 1 II 7 ^Ν Ζ Me. item 59, where the compound is />^0 МеО Т f 1 πσ Н,/* ² Me item 59, where the compound is 3'°Λ·νγ МеО γ Ν Ί πσ ^Η 2 ^Ν ίί Me item 59 , where the compound is \ and ι and ( ^Ν Ζ ^Ν γ^Ν-% Η ₂ Ζ\ Me. p. 59, where the compound is θ^Ν i 3 MeO J ν η io ² Me. p. 59, where the connection is °> ⁿ A ^s ^ meo γ ν η io ^>0° Hol/ ί ² Me item 59, where the connection is - 87 044940 67. The connection according to claim 59, where the connection is 68. The connection according to claim 59, where the connection is claim 59, where the connection is 69. Connection by 70. The connection according to claim 59, where the connection is 71. A compound according to claim 59, wherein the compound is a pharmaceutically acceptable salt of a compound which is 72. A compound according to claim 59, wherein the compound is a pharmaceutically acceptable salt of a compound which is - 88 044940 73. A compound according to claim 59, wherein the compound is a pharmaceutically acceptable salt of a compound which is 74. The compound according to claim 59, where the compound is a pharmaceutically acceptable salt of a compound which is 75. A compound according to claim 59, wherein the compound is a pharmaceutically acceptable salt of a compound which is 76. A compound according to claim 59, wherein the compound is a pharmaceutically acceptable salt of a compound which is 77. A compound according to claim 59, wherein the compound is a pharmaceutically acceptable salt of a compound which is 78. A compound according to claim 59, wherein the compound is a pharmaceutically acceptable salt of a compound which is - 89 044940 79. The compound according to claim 59, where the compound is a pharmaceutically acceptable salt of a compound which is 80. A compound according to claim 59, wherein the compound is a pharmaceutically acceptable salt of a compound which is 81. The compound according to claim 59, where the compound is a pharmaceutically acceptable salt of a compound which is 82. A pharmaceutical composition containing a compound according to claim 59 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. 83. The pharmaceutical composition according to claim 82, wherein the pharmaceutical composition contains the compound according to claim 59 in the form of a free base. 84. The pharmaceutical composition of claim 82, wherein the pharmaceutical composition contains the compound of claim 59 in the form of a pharmaceutically acceptable salt. 85. Use of a compound according to any one of claims 1-28, or 30-57, or 59-81, or a pharmaceutically acceptable salt thereof, for the treatment of a disease treatable by SHP2 inhibition. 86. Use according to claim 85, wherein the disease treatable by SHP2 inhibition is cancer. 87. Use according to claim 86, wherein the cancer is selected from lung cancer, gastric cancer, liver cancer, head and neck cancer, colon cancer, kidney cancer, colorectal cancer, breast cancer, pancreatic cancer, juvenile myelomonocytic leukemia, neuroblastoma, melanoma and acute myeloid leukemia.