RU2819642C1 - Azalactam compounds as inhibitors of hpk1 - Google Patents

Abstract

FIELD: organic chemistry.

SUBSTANCE: group of inventions includes a compound of formula I, pharmaceutically acceptable salts, stereoisomers, individual compounds specified in the claim, a pharmaceutical composition containing them, and methods of treatment. In formula I R¹ is (C₁-C₆)alkyl, -N(R⁵)(R⁶), or (C₃-C₆)cycloalkyl, where said (C₃-C₆)cycloalkyl is substituted with 0 or 1 a substituent which is (C₁-C₆)alkyl or (C₁-C₆)alkoxy; R⁵ and R⁶ each independently represent hydrogen or (C₁-C₆)alkyl, or R⁵ and R⁶ together with nitrogen to which they are attached form (4–8-membered) heterocycloalkyl, which is substituted with 0, 1 or 2 substitutes independently selected from the group consisting of (C₁-C₆)alkyl, halogen(C₁-C₆)alkyl and (C₁-C₆)alkoxy; R² is N(R⁷)(R⁸), each of R⁷ and R⁸ is independently hydrogen or (C₁-C₆)alkyl, which is substituted with 0 or 1 substituent, which is halogen, (C₁-C₆)alkoxy, cyano or hydroxy; R^3a is hydrogen or (C₁-C₃)alkyl substituted with 0 or 1 substituent, which is hydroxy or (C₁-C₃)alkoxy; R^3b is hydrogen or (C₁-C₃)alkyl, provided that R^3a and R^3b are not both H when R⁴ is (R⁴-i); R⁴ is (R⁴-i) or (R⁴-ii); R^4N is (C₁-C₆)alkyl, halogen(C₁-C₆)alkyl or (C₃-C₆)cycloalkyl; R^4C is hydrogen, (C₁-C₆)alkyl, halogen(C₁-C₆)alkyl or (C₁-C₆)alkoxy; R^4D is hydrogen, (C₁-C₆)alkyl, halogen(C₁-C₆)alkyl, halogen(C₁-C₆)alkoxy or (C₃-C₆)cycloalkyl, where said (C₁-C₆)alkyl and halogen(C₁-C₆)alkyl are substituted with 0 or 1 substituent, which is hydroxy or (C₁-C₆)alkoxy; R^4E is hydrogen or (C₁-C₆)alkyl; R^4F is hydrogen, halogen, (C₁-C₆)alkyl or halogen(C₁-C₆)alkyl, where said (C₁-C₆)alkyl and halogen(C₁-C₆)alkyl are substituted with 0 or 1 substitute, which is hydroxy, cyano or (C₁-C₆)alkoxy.

EFFECT: proposed are 2-(pyridin-2-yl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one derivatives of formula I, having inhibitory activity on HPK1 kinase.

21 cl, 8 tbl, 300 ex

Description

Technical field

The present invention relates to compounds of formula I and their pharmaceutically acceptable salts, to pharmaceutical compositions containing such compounds and salts, and to their use. The compounds, salts and compositions of the present invention are HPK1 inhibitors and as such can be used to enhance immune system activation in the treatment or attenuation of abnormal cell proliferative disorders such as cancer and to improve the effectiveness of vaccine therapy.

State of the art

Hematopoietic precursor kinase 1 (HPK1), also known as mitogen-activated protein kinase kinase kinase 1 (MAP4K1), is a member of the Ste20-like family of mammalian serine/threonine kinases that acts through the 7NK and ERK signaling pathways. HPK1 is primarily expressed in hematopoietic organs and cells (e.g., T cells, B cells, and dendritic cells), suggesting a potential involvement of HPK1 in the regulation of signaling in hematopoietic lineages, including lymphocytes. (Shui, et al., “Hematoppietic progenitor kinase 1 negatively regulates T cell receptor signaling and T eel-mediated immune responses,” Nature Immunology 8, 84-91 (2006)).

For example, T cell receptor (TCR) stimulation induces HPK1 tyrosine 379 phosphorylation and its translocation to the plasma membrane. Enzymatic activation of HPK1 is accompanied by phosphorylation of regulatory sites in the HPK1 kinase activation loop. Full activation of HPK1 depends on autophosphorylation of threonine 165 and protein kinase D (PKD) phosphorylation of serine 171 (Arnold et al., “Activation of Hematopoietic Progenitor Kinase 1 Involves Relocation, Autophosphorylation, and Transphosphorylation by Protein Kinase D1.”, Mol Cell Biol 25 (6 ), 2364-83 (2005)). HPK1-mediated phosphorylation of the adapter protein SLP76 ultimately leads to destabilization of the TCR signaling complex, which interferes with and attenuates downstream mitogen-activated protein (MAP) kinase signaling events required for T cell activation and proliferation. (Hernandez, et al., "The kinase activity of hematopoietic progenitor kinase 1 is essential for the regulation of T cell function", Cell Reports 25, (1), 80-94, (2018)). HPK1 kinase has also been shown to negatively regulate T cell signaling through the PGE ₂ receptor in a PKA-dependent manner. In addition, HPK1 kinase has been reported to play a role in: i) activation-induced cell death (AICD) and activation of 7NK; ii) regulating leukocyte function-associated antigen-1 (LFA-1) integrin activation on T cells by directly competing with adhesion and degranulation-promoting adapter protein (ADAP) for binding of the SLP76 SH2 domain; and iii) regulation of activation through nuclear factor κB (NF-κB) signaling through interaction with IKK-α and -β. Studies have also shown that HPK1 negatively regulates the MAP kinase signaling pathway and AP-1 transcription in T cells. (reviewed in Hernandez, et al. 2018).

Research to date on HPK1 kinases suggests that inhibition of HPK1 plays a role in enhancing dendritic and T cell responses and thus enhancing antitumor immunity, viral clearance, and response to vaccine therapy.

Essence

The present invention provides, in part, the compounds of formula I and their pharmaceutically acceptable salts. Such compounds can inhibit HPK1 kinase activity, thereby affecting biological functions. Also provided are pharmaceutical compositions and medicaments containing the compounds or salts of the invention, alone or in combination with additional anticancer therapeutic agents or palliative agents.

The present invention also provides, in part, methods for preparing the compounds, pharmaceutically acceptable salts and compositions of the invention and methods for using the foregoing.

In one embodiment, the invention provides a compound of Formula I:

or a pharmaceutically acceptable salt thereof, where:

R ¹ is (C ₁ -C ₆ )alkyl, halogen (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogen (C ₁ -C ₆ )alkoxy, -N(R ⁵ )(R ⁶ ), or (C ₃ -C ₆ )cycloalkyl, where the specified (C ₁ -C ₆ )alkyl, halogen (C ₁ -C ₆ )alkyl and (C ₃ -C ₆ )cycloalkyl are substituted with 0 or 1 substituent, which represents is hydroxy, cyano, (C ₁ -C ₆ )alkyl or (C ₁ -C ₆ )alkoxy, where:

R ⁵ and R ⁶ are each independently hydrogen or (C ₁ -C ₆ )alkyl substituted with 0, 1 or 2 substituents independently selected from the group consisting of halogen, (C ₁ -C ₆ )alkoxy, cyano and hydroxy, or

R ⁵ and R ⁶ together with the nitrogen to which they are attached form a (4-8 membered) heterocycloalkyl which is substituted with 0, 1 or 2 substituents independently selected from the group consisting of halogen, (C ₁ -C ₆ )alkyl , halogen (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy and halogen (C ₁ -C ₆ )alkoxy;

R ² represents N(R ⁷ )(R ⁸ ), where:

each of R ⁷ and R ⁸ independently represents hydrogen or (C ₁ -C ₆ )alkyl, which is substituted with 0 or 1 substituent representing halogen, (C ₁ -C ₆ )alkoxy, cyano or hydroxy; or

R ⁷ represents hydrogen or (C ₁ -C ₆ )alkyl substituted with 0 or 1 halogen, (C ₁ -C ₆ )alkoxy, cyano or hydroxy substituent; and R ⁸ together with the nitrogen to which it is attached, and together with R ^3a and the carbon to which it is attached, form a (4-6 membered) heterocycloalkyl, which is independently substituted with 0, 1 or 2 substituents selected from halogen, hydroxy , (C ₁ -C ₃ )alkyl, halogen (C ₁ -C ₃ )alkyl, (C ₁ -C ₃ )alkoxy and halogen (C ₁ -C ₆ )alkoxy; or

R ⁷ and R ⁸ together with the nitrogen to which they are attached form a (4-6 membered) heterocycloalkyl which is substituted with 0, 1 or 2 substituents independently selected from halogen, hydroxy, (C ₁ -C ₆ )alkyl, halogen (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy and halogen (C ₁ -C ₆ )alkoxy;

R ^3a represents hydrogen or (C ₁ -C ₃ )alkyl substituted with O or 1 substituent representing hydroxy or (C ₁ -C ₃ )alkoxy;

R ^3b is hydrogen or (C ₁ -C ₃ )alkyl, provided that R ^3a and R ^3b are both not H when R ⁴ is (R ⁴ -i);

R ⁴ is (R ⁴ -i) or (R ⁴ -ii):

Where:

R ^4N represents (C ₁ -C ₆ )alkyl, halogen (C ₁ -C ₆ )alkyl or (C ₃ -C ₆ )cycloalkyl, wherein said (C ₁ -C ₆ )alkyl is substituted with 0 or 1 substituent which represents is hydroxy;

R ^4C is hydrogen, (C ₁ -C ₆ )alkyl, halogen (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogen (C ₁ -C ₆ )alkoxy or (C ₃ -C ₆ )cycloalkyl, wherein said (C ₁ -C ₆ )alkyl and halogen (C ₁ -C ₆ )alkyl are substituted with 0 or 1 substituent which is hydroxy, cyano or (C ₁ -C ₆ )alkoxy;

R ^4D is hydrogen, (C ₁ -C ₆ )alkyl, halogen (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogen (C ₁ -C ₆ )alkoxy or (C ₃ -C ₆ )cycloalkyl, wherein said (C ₁ -C ₆ )alkyl and halogen (C ₁ -C ₆ )alkyl are substituted with 0 or 1 substituent which is hydroxy, cyano or (C ₁ -C ₆ )alkoxy;

R ^4E represents hydrogen, halogen, cyano, hydroxy or (C ₁ -C ₆ )alkyl; And

R ^4F represents hydrogen, halogen, cyano, hydroxy, (C ₁ -C ₆ )alkyl, halogen (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogen (C ₁ -C ₆ )alkoxy or (C ₃ -C ₆ )cycloalkyl, where said (C ₁ -C ₆ )alkyl and halogen (C ₁ -C ₆ )alkyl are substituted with 0 or 1 substituent, which is hydroxy, cyano or (C ₁ -C ₆ )alkoxy .

The invention also relates to therapeutic methods and applications involving the administration of a compound of the invention or a pharmaceutically acceptable salt thereof.

In another embodiment, the invention provides a method of treating abnormal cell growth, particularly cancer, in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof. The compounds of the invention may be administered as single agents or may be administered in combination with other anticancer therapeutic agents, in particular standard of care appropriate for a particular type of cancer.

In another embodiment, the invention provides a method of treating abnormal cell growth, particularly cancer, in a subject in need thereof, comprising administering to the subject an amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, in combination with an amount of an additional anti-cancer therapeutic agent, wherein the amounts together are effective in treating said abnormal cell growth.

In another embodiment, the invention relates to a compound of the invention or a pharmaceutically acceptable salt thereof for use as a drug, in particular a drug for the treatment of cancer.

In another embodiment, the invention provides a compound of the invention, or a pharmaceutically acceptable salt thereof, for use in the treatment of abnormal cell growth, particularly cancer, in a subject.

In another embodiment, the invention provides the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, for the treatment of abnormal cell growth, particularly cancer, in a subject.

In another embodiment, the invention provides a pharmaceutical composition for treating abnormal cell growth in a subject in need thereof, which comprises a compound of the invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

In yet another embodiment, the invention provides the use of a compound of Formula I as described herein, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for the treatment of abnormal cell growth or pathogens in a subject.

In frequent embodiments of the above compounds, methods and uses, the abnormal cell growth is cancer.

In some embodiments, the proposed methods and uses result in one or more of the following effects: (1) inhibition of cancer cell proliferation; (2) inhibition of cancer cell invasiveness; (3) inducing apoptosis of cancer cells; (4) inhibition of cancer cell metastasis; (5) inhibition of angiogenesis; (6) enhancing T cell responses; or (7) enhancement of dendritic and B cell responses; (8) enhancing antitumor activity; (9) strengthening vaccine therapy; and (10) enhancing immune system-mediated removal of pathogens such as viruses, bacteria, or parasites (eg, intestinal worms).

In another embodiment, the invention provides a method of treating HPK1-related disorders and enhancing the immune response in a subject, comprising administering to the subject a compound of the invention or a pharmaceutically acceptable salt thereof in an amount that is effective to treat said disorder or enhance said immune response.

In some embodiments, the methods and uses described herein further comprise administering to the subject an amount of an additional anticancer therapeutic agent, vaccine, antibacterial agent, antiviral agent, or palliative agent that together are effective to treat said abnormal cell growth or pathogen. Each of the embodiments of the compounds of the present invention described below can be combined with one or more other embodiments of the compounds of the present invention described herein without being inconsistent with the embodiment(s) with which it is combined.

It should be understood that both the foregoing general description and the following detailed description are illustrative and explanatory only and do not limit the claimed invention.

In addition, each of the embodiments described below includes pharmaceutically acceptable salts of the compounds of the invention. Accordingly, the phrase “or a pharmaceutically acceptable salt thereof” is intended to refer to all compounds described herein.

Detailed description

Definitions and examples

The present invention may be better understood with reference to the following detailed description of preferred embodiments of the invention and examples included therein. It should be understood that the terminology used herein is for purposes of describing specific embodiments only and is not intended to be limiting. In addition, it should be understood that, unless specifically stated herein, terminology used herein shall have its traditional meaning known in the art.

As used herein, the singular forms "a", "an" and "the" include plural references unless otherwise noted. For example, substituent "a" includes one or more substituents.

The term “about” refers to a relative term indicating the approximation of plus or minus 10% of the nominal value, which, in one embodiment, means plus or minus 5%, in another embodiment, plus or minus 2%. For the scope of this description, this level of approximation is appropriate unless the value is specifically stated as requiring a narrower range.

At various places herein, substituents of the compounds of the invention are described in groups or ranges. In particular, the invention is intended to include each distinct subcombination of members of such groups and ranges. For example, the term "C ₁ - ₆ alkyl" is specifically intended to include C ₁ alkyl (methyl), C ₂ alkyl (ethyl), C ₃ alkyl, C ₄ alkyl, C ₅ alkyl and C ₆ alkyl.

As used herein, the term “HPK1 antagonist” or “HPK1 inhibitor” is a molecule that reduces, inhibits, or otherwise attenuates one or more biological activities of HPK1 (e.g., serine/threonine kinase activity, recruitment to the TCR complex upon TCR activation, interaction with protein binding partner such as SLP76). Antagonism using an HPK1 antagonist does not necessarily indicate complete elimination of HPK1 activity. Instead, activity may decline by a statistically significant amount. For example, a compound of the present invention may reduce HPK1 activity by at least about 2.5% to about 100%, from about 10% to about 90%, from about 20% to about 70%, from about 30% to about 60%, from about 40% to about 50% compared to the corresponding control. In some embodiments, an HPK1 antagonist reduces, inhibits, or otherwise reduces the serine/threonine kinase activity of HPK1. In some of these embodiments, the HPK1 antagonist reduces, inhibits, or otherwise reduces HPK1-mediated phosphorylation of SLP76 and/or Gads. The compounds described herein bind directly to HPK1 and inhibit its kinase activity.

The invention described herein may be practiced in the absence of any element(s) not specifically described herein. Thus, for example, in each instance herein, any of the terms “comprising,” “consisting essentially of,” and “consisting of” may be replaced by any of the other two terms.

The term “(C _x -C _y )alkyl” as used herein refers to a saturated, branched or straight chain alkyl group containing from x to y carbon atoms. For example, "(C ₁ -C ₆ )alkyl" is an alkyl group containing from 1 to 6 carbon atoms and includes, but is not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, vgor-butyl , isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl and n-hexyl. The term "(C ₁ -C ₃ )alkyl" contains from 1 to 3 carbon atoms and is included in "(C ₁ -C ₆ )alkyl".

The term “halogen(C _x -C _y )alkyl” as used herein refers to a (C _x -C _y )alkyl group as defined above, where the alkyl group is replaced by one or more halogen atoms. The typical number of halogen substituents is from 1 to 3 substituents. Typical examples of halogen(C _x -C _y )alkyl include, but are not limited to, fluoromethyl, fluoroethyl, difluoromethyl, difluoroethyl, trifluoromethyl and trifluoroethyl.

The term “(C _x -C _y )alkoxy” as used herein refers to a (C _x -C _y )alkyl group, as defined above, attached to the main molecular group via an oxygen atom. Typical examples of (C ₁ -C ₆ )alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, t-butoxy, pentyloxy and hexyloxy.

The term “halogen(C _x -C _y )alkoxy” as used herein refers to a (C _x -C _y )alkoxy group as defined above, where the alkoxy group is replaced by one or more halogen atoms. The typical number of halogen substituents is from 1 to 3 substituents. Typical examples of halogen(C _x -C _y )alkoxy include, but are not limited to, fluoromethoxy, fluoroethoxy, difluoromethoxy, difluoroethoxy and trifluoromethoxy, trifluoroethoxy.

As used herein, the term “cycloalkyl” refers to a cyclic monovalent hydrocarbon group of the formula _-CnH ( _2n-1 ) containing at least three carbon atoms. "(C ₃ -C _y )cycloalkyl" refers to a cycloalkyl containing from 3 to y carbon atoms. "(C ₃ -C ₆ )cycloalkyl" may be a monocyclic ring, examples of which include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

As used herein, the term “heterocycloalkyl” refers to a cycloalkyl as defined above in which at least one of the ring carbon atoms is replaced by a heteroatom selected from nitrogen, oxygen and sulfur. As used herein, the term “n-membered,” where n is an integer, generally describes the number of ring-forming atoms in a group, where the number of ring-forming atoms is n. The term "(4-6 membered)heterocycloalkyl" means that the heterocycloalkyl substituent contains a total of 4 to 6 ring atoms, at least one of which is a heteroatom. The term "(4-8 membered)heterocycloalkyl" means that the heterocycloalkyl substituent contains a total of 4 to 8 ring atoms, at least one of which is a heteroatom. "(6-membered)heterocycloalkyl" means that the heterocycloalkyl substituent contains a total of 6 ring atoms, at least one of which is a heteroatom. "(5-membered) heterocycloalkyl" means that the heterocycloalkyl substituent contains a total of 5 ring atoms, at least one of which is a heteroatom. The heterocycloalkyl substituent can be attached via a nitrogen atom having the appropriate valency or via any carbon atom in the ring. The heterocycloalkyl group may optionally be substituted with one or more substituents, such as (C ₁ -C ₆ )alkyl, on a nitrogen atom having the appropriate valence or on any available carbon atom.

Examples of heterocycloalkyl rings include, but are not limited to, azetidinyl, tetrahydrofuranyl, tetrahydropyrazolyl, tetrahydrooxazinyl, imidazolidinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxazolidinyl, tetrahydropyranyl, tetrahydrooxazolyl, morpholinyl, and oxetanyl.

"Halo" or "halogen" as used herein refers to a chlorine, fluorine, bromine or iodine atom.

"Hydroxy" or "hydroxyl" as used herein means an -OH group.

"Cyano" as used herein refers to the -CN group, which may also be depicted:

"Patient" or "subject" refers to warm-blooded animals such as, for example, pigs, cows, chickens, horses, guinea pigs, mice, rats, gerbils, cats, rabbits, dogs, monkeys, chimpanzees and humans.

"Pharmaceutically acceptable" means that the substance or composition must be chemically and/or toxicologically compatible with the other ingredients contained in the composition and/or the mammal being treated.

The term “therapeutically effective amount” as used herein refers to that amount of compound administered that will provide some relief from one or more symptoms of the disorder being treated. With respect to the treatment of an HPK1 kinase-mediated disorder (eg, cancer), a therapeutically effective amount refers to that amount that to some extent alleviates (or, for example, eliminates) one or more symptoms associated with the HPK1 kinase-mediated disorder. For example, a therapeutically effective amount refers to that amount which has the effect of (1) reducing tumor size, (2) inhibiting (i.e., to some extent slowing, preferably stopping) tumor metastasis, (3) inhibiting to some extent (i.e., slowing, to some extent, preferably stopping) tumor growth or tumor invasiveness and/or (4) alleviating to some extent (or preferably eliminating) one or more signs or symptoms associated with cancer.

The term “treatment” as used herein, unless otherwise specified, means reversing, alleviating, inhibiting the development of, or preventing the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition. The term “treatment” as used herein, unless otherwise indicated, refers to the act of treating as “treatment” is defined herein. The term "treatment" also includes adjuvant and neoadjuvant treatment of the subject.

"Isomer" means "stereoisomer" and "geometric isomer" as defined below.

"Stereoisomer" refers to compounds that possess one or more chiral centers, each of which can exist in the R or S configuration. Stereoisomers include all diastereomeric, enantiomeric and epimeric forms, as well as racemates and mixtures thereof.

“Geometric isomer” refers to compounds that can exist in cis, trans, anti, entgegen (E) and zusammen (Z) forms, as well as mixtures thereof.

In the present description, the terms "substituent", "radical" and "group" are used interchangeably.

If substituents are described as "independently selected" from a group, each example of substituent is selected independently of any other. Thus, each substituent may be identical to or different from the other substituent(s).

Connections

The compounds of formula I, as described herein, contain an azalactam (2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one) core in which the pyrrolo ring is attached via its nitrogen atom to the pyridine, which substituted with R ⁴ .

In another embodiment, the invention relates to a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R ⁴ is (R ⁴ -i);

R ^4N represents (C ₁ -C ₆ )alkyl or (C ₃ -C ₆ )cycloalkyl; And

R ^4C represents hydrogen or (C ₁ -C ₃ )alkyl; wherein R ¹ , R ² , R ^3a and R ^3b have the meanings defined in any of the embodiments described herein.

In another embodiment, the invention relates to a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R ⁴ is (R ⁴ -i), where R ¹ is N(R ⁵ )(R ⁶ ) or (C ₃ -C ₆ )cycloalkyl, wherein said (C ₃ -C ₆ )cycloalkyl is cyclopropyl and is substituted with 0 or 1 substituent which is (C ₁ -C ₆ )alkyl, wherein said (C ₁ -C ₆ )alkyl is methyl;

R ⁵ and R ⁶ are each independently hydrogen or (C ₁ -C ₃ )alkyl, or

R ⁵ and R ⁶ together with the nitrogen atom to which they are attached form a (5-membered) heterocycloalkyl substituted with 0 or 1 substituent, which is (C ₁ -C ₆ )alkyl;

R ² represents N(R ⁷ )(R ⁸ ), wherein each of R ⁷ and R ⁸ independently represents hydrogen or (C ₁ -C ₆ )alkyl, which represents methyl;

R ^3a is (C ₁ -C ₃ )alkyl substituted with 0 or 1 substituent, which is (C ₁ -C ₃ )alkoxy; And

R ^3b represents hydrogen or (C ₁ -C ₃ )alkyl.

In another embodiment, the invention relates to a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R ⁴ is (R ⁴ -ii);

R ^4D represents hydrogen, (C ₁ -C ₆ )alkyl or halogen(C ₁ -C ₆ )alkyl, wherein said (C ₁ -C ₆ )alkyl is substituted with 0 or 1 substituent which is hydroxy;

R ^4E represents hydrogen; And

R ^4F represents hydrogen, (C ₁ -C ₆ )alkyl or halogen(C ₁ -C ₆ )alkyl, wherein said (C ₁ -C ₆ )alkyl is substituted with 0 or 1 substituent which is hydroxy; and wherein R ¹ , R ² , R ^3a and R ^3b are as defined in any of the embodiments described herein.

In another embodiment, the invention relates to a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R ⁴ is (R ⁴ -ii), and where R ¹ is -N(R ⁵ )(R ⁶ ) or (C ₃ - C ₆ )cycloalkyl, wherein said (C ₃ -C ₆ )cycloalkyl is cyclopropyl and is substituted with 0 or 1 substituent which is (C ₁ -C ₆ )alkyl, wherein said (C ₁ -C ₆ )alkyl is methyl ;

R ⁵ and R ⁶ are each independently hydrogen or (C ₁ -C ₃ )alkyl, or

R ⁵ and R ⁶ together with the nitrogen atom to which they are attached form a (5-membered) heterocycloalkyl substituted with 0 or 1 substituent, which is (C ₁ -C ₆ )alkyl;

R ² represents N(R ⁷ )(R ⁸ ), wherein each of R ⁷ and R ⁸ independently represents hydrogen or (C ₁ -C ₆ )alkyl, which represents methyl;

R ^3a represents hydrogen or (C ₁ -C ₃ )alkyl substituted with O or 1 substituent, which is (C ₁ -C ₃ )alkoxy; And

R ^3b represents hydrogen or (C ₁ -C ₃ )alkyl.

In another embodiment, the invention relates to a compound of Formula I or a pharmaceutically acceptable salt thereof, wherein R ¹ is 2-methylpyrrolidin-1-yl or 2(R)-methylpyrrolidin-1-yl; and wherein R ² , R ^3a , R ^3b and R ⁴ are as defined in any of the embodiments described herein.

In another embodiment, the invention provides a compound of Formula I or a pharmaceutically acceptable salt thereof, wherein R ² is N(R ⁷ )(R ⁸ ), wherein R ⁷ and R ⁸ are each hydrogen, and where R ¹ , R ^3a , R ^3b and R ⁴ have the meanings defined in any of the embodiments described herein.

In another embodiment, the invention relates to a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R ^3a is (C ₁ -C ₃ )alkyl substituted with 0 or 1 substituent which is (C ₁ -C ₃ )alkoxy; and wherein R ¹ , R ² , R ^3b and R ⁴ are as defined in any of the embodiments described herein.

In another embodiment, the invention relates to a compound of Formula I or a pharmaceutically acceptable salt thereof, wherein R ^3b is (C ₁ -C ₃ )alkyl; and R ¹ , R ² , R ^3a and R ⁴ have the meanings defined in any of the embodiments described herein.

In another embodiment, the invention provides a compound of Formula I or a pharmaceutically acceptable salt thereof, wherein R ² is N(R ⁷ )(R ⁸ ), wherein R ⁷ and R ⁸ are each hydrogen; and wherein R ¹ , R ^3a , R ^3b and R ⁴ are as defined in any of the embodiments described herein.

In another embodiment, the invention relates to a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R ² is N(R ⁷ )(R ⁸ ), wherein R ⁷ is hydrogen; and R ⁸ represents (C ₁ -C ₃ )alkyl; and wherein R ¹ , R ^3a , R ^3b and R ⁴ are as defined in any of the embodiments described herein.

In another embodiment, the invention provides a compound of formula IA (a compound of formula I wherein R ⁴ is (R ⁴ -i):

or a pharmaceutically acceptable salt thereof, where:

R ¹ represents -N(R ⁵ )(R ⁶ ) or (C ₃ -C ₆ )cycloalkyl, wherein said (C ₃ -C ₆ )cycloalkyl represents cyclopropyl and is substituted with 0 or 1 substituent, that is, (C ₁ - C ₆ )alkyl, wherein said (C ₁ -C ₆ )alkyl is methyl;

R ⁵ and R ⁶ are each independently hydrogen or (C ₁ -C ₃ )alkyl, or

R ⁵ and R ⁶ together with the nitrogen atom to which they are attached form a (5-membered) heterocycloalkyl substituted with 0 or 1 substituent, which is (C ₁ -C ₆ )alkyl;

R ² is N(R ⁷ )(R ⁸ ), wherein R ⁷ and R ⁸ are each independently hydrogen or (C ₁ -C ₆ )alkyl, which is methyl;

R ^3a is (C ₁ -C ₃ )alkyl substituted with 0 or 1 substituent, which is (C ₁ -C ₃ )alkoxy;

R ^3b represents hydrogen or (C ₁ -C ₃ )alkyl;

R ^4N represents (C ₁ -C ₆ )alkyl or (C ₃ -C ₆ )cycloalkyl; And

R ^4C represents hydrogen or (C ₁ -C ₃ )alkyl.

In another embodiment, the invention relates to a compound of formula IA or a pharmaceutically acceptable salt thereof, wherein R ^3a is (C ₁ -C ₃ )alkyl substituted with 0 or 1 substituent which is (C ₁ -C ₃ )alkoxy; And

R ^3b represents (C ₁ -C ₃ )alkyl.

In some embodiments, when R ⁴ is (R ⁴ -i), the compound of Formula IA has absolute stereochemistry as shown in Formula IAi or IA-ii:

or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ² , R ^3a , R ^3b and R ^4C are defined in any embodiment of Formula IA.

In another embodiment, the invention relates to a compound of formula IB (a compound of formula I, wherein R ⁴ is (R ⁴ -ii):

or a pharmaceutically acceptable salt thereof, where:

R ¹ is (C ₁ -C ₆ )alkyl, halogen (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogen (C ₁ -C ₆ )alkoxy, -N(R ⁵ )(R ⁶ ), or (C ₃ -C ₆ )cycloalkyl, where the specified (C ₁ -C ₆ )alkyl, halogen (C ₁ -C ₆ )alkyl and (C ₃ -C ₆ )cycloalkyl are substituted with 0 or 1 substituent, which represents is hydroxy, cyano, (C ₁ -C ₆ )alkyl or (C ₁ -C ₆ )alkoxy, where:

R ⁵ and R ⁶ are each independently hydrogen or (C ₁ -C ₆ )alkyl substituted with 0, 1 or 2 substituents independently selected from the group consisting of halogen, (C ₁ -C ₆ )alkoxy, cyano and hydroxy, or

R ⁵ and R ⁶ together with the nitrogen to which they are attached form a (4-8 membered) heterocycloalkyl which is substituted with 0, 1 or 2 substituents independently selected from the group consisting of halogen, (C ₁ -C ₆ )alkyl , halogen (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy and halogen (C ₁ -C ₆ )alkoxy;

R ² represents N(R ⁷ )(R ⁸ ), where:

each of R ⁷ and R ⁸ independently represents hydrogen or (C ₁ -C ₆ )alkyl, which is substituted with 0 or 1 substituent representing halogen, (C ₁ -C ₆ )alkoxy, cyano or hydroxy; or

R ⁷ represents hydrogen or (C ₁ -C ₆ )alkyl substituted with O or 1 substituent representing halogen, (C ₁ -C ₆ )alkoxy, cyano or hydroxy; and R ⁸ together with the nitrogen to which it is attached, and together with R ^3a and the carbon to which it is attached, form a (4-6 membered) heterocycloalkyl, which is independently substituted with 0, 1 or 2 substituents selected from halogen, hydroxy , (C ₁ -C ₃ )alkyl, halogen (C ₁ -C ₃ )alkyl, (C ₁ -C ₃ )alkoxy or halogen(C ₁ -C ₆ )alkoxy; or

R ⁷ and R ⁸ together with the nitrogen to which they are attached form a (4-6 membered) heterocycloalkyl which is substituted with 0, 1 or 2 substituents independently selected from halogen, hydroxy, (C ₁ -C ₆ )alkyl, halogen (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy and halogen(C ₁ -C ₆ )alkoxy;

R ^3a represents hydrogen or (C ₁ -C ₃ )alkyl substituted with O or 1 substituent representing hydroxy or (C ₁ -C ₃ )alkoxy;

R ^3b represents hydrogen or (C ₁ -C ₃ )alkyl;

R ^4D is hydrogen, (C ₁ -C ₆ )alkyl, halogen(C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogen(C ₁ -C ₆ )alkoxy or (C ₃ -C ₆ )cycloalkyl, wherein said (C ₁ -C ₆ )alkyl and halogen (C ₁ -C ₆ )alkyl are substituted with 0 or 1 substituent which is hydroxy, cyano or (C ₁ -C ₆ )alkoxy;

R ^4E represents hydrogen, halogen, cyano, hydroxy or (C ₁ -C ₆ )alkyl; And

R ^4F is hydrogen, halogen, cyano, hydroxy, (C ₁ -C ₆ )alkyl, halogen (C ₁ -C ₆ )alkyl, (C ₁ -C ₆ )alkoxy, halogen (C ₁ -C ₆ )alkoxy or (C ₃ -C ₆ )cycloalkyl, where said (C ₁ -C ₆ )alkyl and halogen(C ₁ -C ₆ )alkyl are substituted with 0 or 1 substituent, which is hydroxy, cyano or (C ₁ -C ₆ )alkoxy ; and wherein R ² has the meaning defined in any of the embodiments described herein.

In another embodiment, the invention relates to a compound of formula I-B or a pharmaceutically acceptable salt thereof, wherein:

R ¹ represents -N(R ⁵ )(R ⁶ ) or (C ₃ -C ₆ )cycloalkyl, wherein said (C ₃ -C ₆ )cycloalkyl is substituted with 0 or 1 substituent which is (C ₁ -C ₃ ) alkyl, where:

R ⁵ and R ⁶ are each independently hydrogen or (C ₁ -C ₆ )alkyl, or

R ⁵ and R ⁶ together with the nitrogen atom to which they are attached form a (4-8 membered) heterocycloalkyl substituted with 0, 1 or 2 substituents, which is (C ₁ -C ₆ )alkyl;

R ² represents N(R ⁷ )(R ⁸ ), wherein each of R ⁷ and R ⁸ independently represents hydrogen or (C ₁ -C ₆ )alkyl;

R ^3a represents hydrogen or (C ₁ -C ₃ )alkyl substituted with 0 or 1 substituent, which is (C ₁ -C ₃ )alkoxy;

R ^3b represents hydrogen or (C ₁ -C ₃ )alkyl;

R ^4D represents hydrogen, (C ₁ -C ₃ )alkyl or halogen (C ₁ -C ₃ )alkyl, wherein said (C ₁ -C ₃ )alkyl is substituted with 0 or 1 substituent which is hydroxy, cyano or (C ₁ -C ₃ )alkoxy;

R ^4E represents hydrogen; And

R ^4F represents hydrogen, (C ₁ -C ₃ )alkyl or halogen(C ₁ -C ₃ )alkyl, wherein said (C ₁ -C ₃ )alkyl is substituted with 0 or 1 substituent which is hydroxy.

In another embodiment, the invention provides a compound of Formula IB or a pharmaceutically acceptable salt thereof, wherein R ^3b is hydrogen or (C ₁ -C ₃ )alkyl.

In another embodiment, the invention relates to a compound of formula IB or a pharmaceutically acceptable salt thereof, wherein R ^4D is hydrogen, (C ₁ -C ₃ )alkyl or halogen(C ₁ -C ₃ )alkyl, wherein said (C ₁ -C ) ₃ ) alkyl is substituted with 0 or 1 substituent, which is hydroxy; and wherein R ¹ , R ^3a , R ^3b , R ^4E and R ^4F are as defined in any of the embodiments described herein.

In another embodiment, the invention relates to a compound of formula IB or a pharmaceutically acceptable salt thereof, wherein R ^3a is (C ₁ -C ₃ )alkyl substituted with 0 or 1 substituent which is (C ₁ -C ₃ )alkoxy; And

R ^3b represents (C ₁ -C ₃ )alkyl; and wherein R ¹ , R ² , R ^4D , R ^4E and R ^4F are as defined in any of the embodiments described herein.

In another embodiment, the invention provides a compound of formula I-B or a pharmaceutically acceptable salt thereof, wherein:

R ¹ represents -N(R ⁵ )(R ⁶ ) or (C ₃ -C ₆ )cycloalkyl, where (C ₃ -C ₆ )cycloalkyl is substituted with 0 or 1 (C ₁ -C ₃ )alkyl;

R ⁵ and R ⁶ are each independently selected from the group consisting of (C ₁ -C ₆ )alkyl, or R ⁵ and R ⁶ together with the nitrogen atom to which they are attached form a (5-membered) heterocycloalkyl which is substituted with 0 or 1 (C ₁ -C ₆ )alkyl;

R ² represents N(R ⁷ )(R ⁸ ), wherein each of R ⁷ and R ⁸ represents hydrogen;

R ^3a represents hydrogen or (C ₁ -C ₃ )alkyl, wherein said (C ₁ -C ₃ )alkyl is substituted with 0 or 1 (C ₁ -C ₃ )alkoxy group;

R ^3b represents hydrogen or (C ₁ -C ₃ )alkyl;

R ^4D represents hydrogen or (C ₁ -C ₃ )alkyl, where (C ₁ -C ₃ )alkyl is substituted with 0 to 1 substituent selected from OH and F;

R ^4E represents hydrogen; And

R ^4F represents hydrogen or (C ₁ -C ₃ )alkyl.

In another embodiment, the invention relates to a compound of formula I-B or a pharmaceutically acceptable salt thereof, wherein:

R ¹ represents -N(R ⁵ )(R ⁶ ) or cyclopropyl substituted with 0 or 1 methyl substituent;

R ⁵ and R ⁶ are each independently hydrogen or (C ₁ -C ₃ )alkyl, or

R ⁵ and R ⁶ together with the nitrogen atom to which they are attached form a (5-membered) heterocycloalkyl substituted with 0 or 1 substituent, which is methyl;

R ² is N(R ⁷ )(R ⁸ ), wherein R ⁷ and R ⁸ are each independently hydrogen or methyl;

R ^3a is (C ₁ -C ₃ )alkyl substituted with 0 or 1 substituent, which is (C ₁ -C ₃ )alkoxy; And

R ^3b represents hydrogen or (C ₁ -C ₃ )alkyl; and wherein R ^4D , R ^4E and R ^4F have the meanings defined in any of the embodiments described herein.

In another embodiment, the invention relates to a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R ⁴ is (R ⁴ -ii);

R ^4D represents hydrogen, (C ₁ -C ₆ )alkyl or halogen (C ₁ -C ₆ )alkyl, wherein said (C ₁ -C ₆ )alkyl is substituted with 0 or 1 substituent which is hydroxy;

R ^4E represents hydrogen; And

R ^4F represents hydrogen, (C ₁ -C ₆ )alkyl or halogen(C ₁ -C ₆ )alkyl, wherein said (C ₁ -C ₆ )alkyl is substituted with 0 or 1 substituent which is hydroxy; and wherein R ¹ , R ² , R ^3a and R ^3b are as defined in any of the embodiments described herein.

In another embodiment, the invention relates to a compound of formula I or a pharmaceutically acceptable salt thereof, wherein R ⁴ is (R ⁴ -ii);

R ^4D is hydrogen, methyl, fluoromethyl, hydroxymethyl or ethyl;

R ^4E represents hydrogen; And

R ^4F represents hydrogen; and wherein R ¹ , R ² , R ^3a and R ^3b are as defined in any of the embodiments described herein.

In another embodiment, the invention relates to compounds of formula I or pharmaceutically acceptable salts thereof having specific stereochemistry with respect to the orientation of R ² , R ^3a and R ^3b when R ^3a is different from R ^3b . For example, when R ^3b is hydrogen (R ^3b is absent), the compounds of Formula I will look like this:

where R ¹ , R ^3a and R ⁴ have the meanings defined in any of the embodiments described herein.

In another embodiment, the invention relates to a compound of formula I(R) or formula I(S) or a pharmaceutically acceptable salt thereof, wherein:

R ¹ is -N(R ⁵ )(R ⁶ ) or cyclopropyl substituted with one substituent that is methyl;

R ⁵ and R ⁶ are each independently hydrogen or (C ₁ -C ₃ )alkyl, or

R ⁵ and R ⁶ together with the nitrogen atom to which they are attached form a (5-membered) heterocycloalkyl substituted with 0 or 1 substituent, which is methyl;

R ² is N(R ⁷ )(R ⁸ ), wherein R ⁷ and R ⁸ are each independently hydrogen or methyl;

R ^3a is (C ₁ -C ₃ )alkyl substituted with 0 or 1 substituent, which is (C ₁ -C ₃ )alkoxy; R ^3b represents hydrogen; R ⁴ is (R ⁴ -ii);

R ^4D is hydrogen, methyl, fluoromethyl, hydroxymethyl or ethyl;

R ^4E represents hydrogen; And

R ^4F represents hydrogen or methyl.

In yet another embodiment, the invention relates to compounds of formula I (R) or pharmaceutically acceptable salts thereof, wherein R ^3b is hydrogen (R ^3b is absent):

and wherein R ¹ , R ^3a and R ⁴ have the meanings defined in any of the embodiments described herein.

In yet another embodiment, the invention relates to compounds of formula I (S) or pharmaceutically acceptable salts thereof, wherein R ^3b is hydrogen (R ^3b is absent):

wherein R ¹ , R ^3a and R ⁴ have the meanings defined in any of the embodiments described herein.

In another embodiment, the invention provides a compound of formula IB or a pharmaceutically acceptable salt thereof, wherein R ⁴ is 5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole -3-yl, (5R)-5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl, (5S)-5-methyl- 6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl, 5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][ 1,2,4]triazol-3-yl, (5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl or ( 5R)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl, wherein said methyl is substituted with 0 or 1 substituent which is F or OH, and wherein R ¹ , R ² , R ^3a and R ^3b are as defined in any of the embodiments described herein.

In another embodiment, the invention provides a compound of formula IB or a pharmaceutically acceptable salt thereof, wherein R ⁴ is (5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2 ,4]triazol-3-yl or (5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl, wherein said methyl is substituted 0 or 1 substituent that is F or OH, and wherein R ¹ , R ² , R ^3a and R ^3b are as defined in any of the embodiments described herein.

Each of the embodiments described herein with respect to Formula I also applies to compounds of Formulas I-A-i, I-A-ii, I-B-i and I-B-ii.

In another embodiment, the invention provides a compound wherein the compound is: 4-[1-aminopropyl]-2-{6-[5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1 ,2,4]triazol-3-yl]pyridin-2-yl}-6-[2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1- He;

4-[1-aminoethyl]-2-{6-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl]pyridin-2- yl}-6-[2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one;

4-[1-aminoethyl]-2-{6-[5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl]pyridin-2 -yl}-6-[2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one;

4-[1-aminoethyl]-2-{6-[5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl]pyridin-2 -yl}-6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one;

4-[1-aminopropyl]-2-{3-[5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl]phenyl}- 6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one; or

4-[1-amino-2-methoxyethyl]-6-(1-methylcyclopropyl)-2-{6-[5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2 ,4]triazol-3-yl]pyridin-2-yl}-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one; or a pharmaceutically acceptable salt thereof.

For example, the compound of the invention is a compound which is 4-[(1R)-1-aminopropyl]-2-{6-[(5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2. 1-c][1,2,4]triazol-3-yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[ 3,4-c]pyridin-1-one;

4-[{1R)-1-aminoethyl]-2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrole o[2,1-c][1,2,4] triazol-3-yl]pyridin-2-yl}-6-[{2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one ;

4-[{1S)-1-aminoethyl]-2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole -3-yl]pyridin-2-yl}-6-[{2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one;

4-[(1R)-1-aminoethyl]-2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole -3-yl]pyridin-2-yl}-6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one;

4-[(1R)-1-aminopropyl]-2-{3-[(5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole -3-yl]phenyl}-6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one;

4-[{1R)-1-amino-2-methoxyethyl]-6-(1-methylcyclopropyl)-2-{6-[(5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2, 1-c][1,2,4]triazol-3-yl]pyridin-2-yl}-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one; or

4-[{1S)-1-amino-2-methoxyethyl]-6-(1-methylcyclopropyl)-2-{6-[(5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2, 1-c][1,2,4]triazol-3-yl]pyridin-2-yl}-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one; or a pharmaceutically acceptable salt thereof.

In another embodiment, the invention relates to compounds of formula I or pharmaceutically acceptable salts thereof, wherein the compounds are compounds of formula 1(R) or compounds of formula I(S), having specific stereochemistry with respect to the orientation of R ² , R ^3a and R ^3b when R ^3a differs from R ^3b .

In another embodiment, the invention relates to a compound that is 4-[(1R)-1-aminopropyl]-2-{6-[(5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2 ,1-c][1,2,4]triazol-3-yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo [3,4-c]pyridin-1-one; or a pharmaceutically acceptable salt thereof.

In another embodiment, the invention relates to a compound that is 4-[(1R)-1-aminoethyl]-2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2 ,1-c][1,2,4]triazol-3-yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo [3,4-c]pyridin-1-one; or a pharmaceutically acceptable salt thereof.

In another embodiment, the invention relates to compounds that are 4-[(1S)-1-aminoethyl]-2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2 ,1-c][1,2,4]triazol-3-yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo [3,4-c]pyridin-1-one; or a pharmaceutically acceptable salt thereof.

In another aspect, the invention provides a compound selected from the group consisting of the compounds exemplified herein, or a pharmaceutically acceptable salt thereof.

The compounds of the invention are selective for HPK1 kinase.

"Pharmaceutical composition" refers to a mixture of one or more compounds described herein, or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof, as an active ingredient, and at least one pharmaceutically acceptable carrier or excipient. In some embodiments,

the pharmaceutical composition contains two or more pharmaceutically acceptable carriers and/or excipients.

In some embodiments, the pharmaceutical composition further comprises at least one additional anticancer therapeutic agent or palliative agent. In some such embodiments, the at least one additional agent is an anticancer therapeutic agent, as described below. In some such embodiments, the combination offers additive, greater than additive, or synergistic anticancer effects.

In one embodiment, the invention provides a method of treating abnormal cell growth in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the invention or a pharmaceutically acceptable salt thereof.

In another embodiment, the invention provides a method of treating abnormal cell growth in a subject in need thereof, comprising administering to the subject an amount of a compound of the invention or a pharmaceutically acceptable salt thereof in combination with an amount of an additional therapeutic agent (eg, an anticancer therapeutic agent), wherein the amount in the aggregate is effective in treating said abnormal cell growth.

In frequent embodiments of the methods provided herein, the abnormal cell growth is cancer. The compounds of the invention may be administered as single agents or may be administered in combination with other anticancer therapeutic agents, in particular standard of care, appropriate for a particular type of cancer.

In some embodiments, the proposed methods result in one or more of the following effects: (1) inhibition of cancer cell proliferation; (2) inhibition of cancer cell invasiveness; (3) inducing apoptosis of cancer cells; (4) inhibition of cancer cell metastasis; (5) inhibition of angiogenesis; (6) enhancing T cell responses; (7) enhanced dendritic and B cell responses; (8) enhancing antitumor activity; (9) strengthening vaccine therapy; and (10) enhancing immune system-mediated removal of pathogens such as viruses, bacteria, worms.

In another aspect, the invention provides a method of treating a disorder mediated by HPK1 kinase activity in a subject, such as certain types of cancer, comprising administering to the subject a compound of the invention or a pharmaceutically acceptable salt thereof in an amount that is effective to treat the disorder.

Unless otherwise indicated, all references herein to the compounds of the invention include references to their salts, solvates, hydrates and complexes, as well as their solvates, hydrates and salt complexes, including polymorphs, stereoisomers and isotopically labeled variants thereof.

The compounds of the invention may exist in the form of pharmaceutically acceptable salts, such as, for example, the acid addition salts and base addition salts of the compounds of Formula I provided herein. As used herein, the term “pharmaceutically acceptable salt” refers to those salts that retain the biological effectiveness and properties of the parent compound. The phrase "pharmaceutically acceptable salt(s)" as used herein, unless otherwise indicated, includes salts of acidic or basic groups that may be present in the compounds of Formula I described herein.

For example, the compounds of the invention, which are basic in nature, are capable of forming a wide range of salts with various inorganic and organic acids. Although such salts should be pharmaceutically acceptable for administration to animals, in practice it is often desirable to first isolate the compound of the present invention from the reaction mixture as a pharmaceutically unacceptable salt, and then simply convert the latter back to the free base compound by treatment with an alkaline reagent, and then convert the latter free base into a pharmaceutically acceptable acid addition salt. Acid addition salts of the base compounds of the present invention can be prepared by treating the base compound with a substantially equivalent amount of a selected mineral or organic acid in an aqueous solvent or a suitable organic solvent such as methanol or ethanol. After evaporation of the solvent, the desired solid salt is obtained. The desired acid salt can also be precipitated from a solution of the free base in an organic solvent by adding a suitable mineral or organic acid to the solution.

Acids that can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds are those that form non-toxic acid addition salts, i.e. salts containing pharmacologically acceptable anions such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, acid citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate , gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate [i.e. 1,1'-methylene-bis(2-hydroxy-3-naphthoate)].

Examples of salts include, but are not limited to, acetate, acrylate, benzenesulfonate, benzoate (for example, chlorobenzoate, methyl benzoate, dinitrobenzoate, hydroxybenzoate and methoxybenzoate), bicarbonate, bisulfate, bisulfite, bitartrate, borate, bromide, butine-1,4-dioate, calcium edetate, camsylate, carbonate, chloride, caproate, caprylate, clavulanate, citrate, decanoate, dihydrochloride, dihydrogen phosphate, edetate, edisilate, estolate, esylate, ethyl succinate, formate, fumarate, gluceptate, gluconate, glutamate, glycolate, glycolylarsanilate, heptanoate, hexine -1,6-dioate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, γ-hydroxybutyrate, iodide, isobutyrate, isothionate, lactate, lactobionate, laurate, malate, maleate, malonate, mandelate, mesylate, metaphosphate, methanesulfonate, methyl sulfate, monohydrogen phosphate, mucate , napsylate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, nitrate, oleate, oxalate, pamoate (embonate), palmitate, pantothenate, phenylacetates, phenylbutyrate, phenylpropionate, phthalate, phosphate/diphosphate, polygalacturonate, propanesulfonate, propionate, propiolate, pyrophosphate, pyrosulfate, salicylate, stearate, subacetate, suberate, succinate, sulfate, sulfonate, sulfite, tannate, tartrate, theoclate, tosylate, triethiododo and valerate.

Illustrative examples of suitable salts include organic salts derived from amino acids such as glycine and arginine, ammonia, primary, secondary and tertiary amines and cyclic amines such as piperidine, morpholine and piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium.

The compounds of the invention, which include a basic group such as an amino group, can form pharmaceutically acceptable salts with various amino acids in addition to the acids mentioned above.

Those compounds of the invention which are acidic in nature are capable of forming basic salts with various pharmacologically acceptable cations. Examples of such salts include alkali or alkaline earth metal salts and, in particular, sodium and potassium salts. All these salts are obtained by conventional methods. The chemical bases that are used as reagents to prepare the pharmaceutically acceptable base salts of the present invention are bases that form non-toxic basic salts with acidic compounds herein. These salts can be prepared by any suitable method, for example by treating the free acid with an inorganic or organic base such as an amine (primary, secondary or tertiary), an alkali metal hydroxide or an alkaline earth metal hydroxide or the like. These salts can also be prepared by treating the corresponding acidic compounds with an aqueous solution containing the desired pharmacologically acceptable cations and then evaporating the resulting solution to dryness, preferably under reduced pressure. Alternatively, they can also be prepared by mixing lower alkanol solutions of acidic compounds and the desired alkali metal alkoxide together and then evaporating the resulting solution to dryness in the same manner as before. In any case, it is preferable to use stoichiometric amounts of reactants to ensure completeness of the reaction and maximum yield of the desired final product.

Chemical bases that can be used as reagents to prepare pharmaceutically acceptable base salts of the compounds of the invention that are acidic in nature are those that form non-toxic base salts with such compounds. Such non-toxic base salts include, but are not limited to, addition salts derived from pharmacologically acceptable cations such as alkali metal cations (eg, potassium and sodium) and alkaline earth metal cations (eg, calcium and magnesium), ammonium or water-soluble amine, such as N-methylglucamine (meglumine), lower alkanolammonium and other basic salts of pharmaceutically acceptable organic amines.

Hemisalts of acids and bases can also be formed, for example, themesulfate and hemicalcium salts.

For a review of suitable salts, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use by Stahl and Wermuth (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds of the invention are known to those skilled in the art.

The salts of the present invention can be prepared by methods known to those skilled in the art. A pharmaceutically acceptable salt of the compounds of the invention can be readily prepared by mixing together solutions of the compound and the desired acid or base, as appropriate. The salt may precipitate out of solution and be collected by filtration, or may be isolated by evaporation of the solvent. The degree of ionization in salt can vary from completely ionized to almost no ionization.

Those skilled in the art will appreciate that the compounds of the invention in free base form having basic functionality can be converted into acid addition salts by treatment with a stoichiometric excess of the appropriate acid. The acid addition salts of the compounds of the invention can be reconverted to the corresponding free base by treatment with a stoichiometric excess of a suitable base, such as potassium carbonate or sodium hydroxide, typically in the presence of an aqueous solvent and at a temperature between about 0°C and 100°C. The free base form can be isolated by conventional methods such as organic solvent extraction. In addition, the acid addition salts of the compounds of the present invention can be replaced by taking advantage of different salt solubility, volatility or acidity of the acids, or by treating with an ion exchange resin with an appropriate content. For example, the exchange may be affected by the reaction of a salt of the compounds of the invention with a small stoichiometric excess of an acid with a lower pK than the acidic component of the parent salt. This transformation is usually carried out at a temperature from about 0°C to the boiling point of the solvent used as the medium for the procedure. Similar exchanges are possible with base addition salts, usually via the free base form.

The compounds of the invention may exist in both non-solvated and solvated forms. When the solvent or water is tightly bound, the complex will have a well-defined stoichiometry that is independent of humidity. However, when the solvent or water is weakly bound, as in channel solvates and hygroscopic compounds, the water/solvent content will depend on the humidity and drying conditions. In such cases, non-stoichiometry will be the norm. The term "solvate" is used herein to describe a molecular complex containing a compound of the invention and one or more molecules of a pharmaceutically acceptable solvent, such as ethanol. The term "hydrate" is used when the solvent is water. Pharmaceutically acceptable solvates of the invention include hydrates and solvates in which the crystallization solvent may be isotopically substituted, for example D ₂ O, d ₆ -acetone, d ₆ -DMSO.

Also included within the scope of the invention are complexes such as clathrates, drug-host inclusion complexes in which, unlike the aforementioned solvates, the drug and host are present in stoichiometric or non-stoichiometric amounts. Also included are drug complexes containing two or more organic and/or inorganic components, which may be present in stoichiometric or non-stoichiometric amounts. The resulting complexes can be ionized, partially ionized, or not

ionized. For a review of such complexes, see 7 Pharm Sci, 64 (8), 1269-1288, Haleblian (August 1975), the description of which is incorporated herein by reference in its entirety.

The invention also relates to prodrugs of the compounds of formula I presented herein. Thus, certain derivatives of the compounds of the invention, which may have little or no pharmacological activity when administered to a patient, can be converted into the compounds of the invention, for example, by hydrolytic cleavage. Such derivatives are called "prodrugs". Additional information on the use of prodrugs can be found in Pro-Drugs as Novel Delivery Systems, Vol.14, ACS Symposium Series (T Higuchi and W Stella) and "Bioreversible Carriers in Drug Design", Pergamon Press, 1987 (ed. E In Roche, American Pharmaceutical Association), the description of which is incorporated herein by reference in its entirety.

Prodrugs according to the invention can be prepared, for example, by replacing the corresponding functional groups present in the compounds of the invention with certain groups known to those skilled in the art as "pro-groups", as described, for example, in "Design of Prodrugs" by H Bundgaard (Elsevier, 1985), the description of which is incorporated herein by reference in its entirety.

Some non-limiting examples of prodrugs of the invention include:

(i) if the compound contains a carboxylic acid functional group (-COOH), its ester, for example replacing hydrogen with (C ₁ -C ₆ )alkyl;

(ii) if the compound contains an alcohol functional group (-OH), its ether, for example, replacement of hydrogen by (C ₁ -C ₆ )alkanoyloxymethyl or phosphate ester group; And

(iii) if the compound contains a primary or secondary amino group (-NH ₂ or -NHR, where R≠H), its amide, for example, the replacement of one or both hydrogen atoms with a suitable metabolically labile group such as amide, carbamate, urea, phosphonate, sulfonate, etc.

Additional examples of substituent groups in accordance with the above examples and examples of other types of prodrugs can be found in the above references.

Finally, some compounds of the invention may themselves act as prodrugs of other compounds of the invention.

Also included within the scope of the invention are metabolites of compounds of formula I as described herein, i.e. compounds formed in vivo upon administration of a drug.

The compounds of formula I presented herein may have asymmetric carbon atoms. The carbon-carbon bonds of compounds of the invention may be represented herein by a solid line ( ), solid wedge ( ) or dotted wedge ( ). Using a solid line to indicate bonds to asymmetric carbon atoms means that all possible stereoisomers (eg, specific enantiomers, racemic mixtures, etc.) on that carbon atom are included. The use of a solid or dotted wedge to depict bonds to asymmetric carbon atoms means that only the stereoisomer shown is intended to be included. It is possible that the compounds of the invention may contain more than one asymmetric carbon atom. In these compounds, the use of a solid line to indicate bonds with asymmetric carbon atoms means that all possible stereoisomers must be included and the stereocenter is attached. For example, unless otherwise indicated, it is intended that the compounds of the invention may exist as enantiomers and diastereomers or as racemates and mixtures thereof. The use of a solid line to represent bonds to one or more asymmetric carbon atoms in a compound of the invention, and the use of a solid or dotted wedge to depict bonds to other asymmetric carbon atoms in the same compound, means that a mixture of diastereomers is present.

The compounds of the invention which have chiral centers may exist as stereoisomers such as racemates, enantiomers or diastereomers.

Stereoisomers of the compounds of Formula I herein may include cis and trans isomers, optical isomers such as (R) and (S) enantiomers, diastereomers, geometric isomers, rotational isomers, atropisomers, conformational isomers and tautomers of the compounds of the invention, including the compounds , exhibiting more than one type of isomerism; and mixtures thereof (such as racemates and diastereomeric pairs).

Also included are acid addition salts or base addition salts in which the counterion is optically active, such as d-lactate or l-lysine, or racemic, such as d-tartrate or d-arginine.

When any racemate crystallizes, two different types of crystals are possible. The first type is the racemic compound mentioned above (true racemate), which produces one homogeneous crystal form containing both enantiomers in equimolar amounts. The second type is a racemic mixture or conglomerate, where the two crystal forms are formed in equimolar quantities, each containing one enantiomer.

The compounds of the invention may exhibit the phenomenon of tautomerism and structural isomerism. For example, compounds can exist in several tautomeric forms, including enol and imine forms, as well as keto and enamine forms, and geometric isomers and mixtures thereof. All such tautomeric forms are included within the scope of the compounds of the invention. Tautomers exist as mixtures of the tautomeric set in solution. In solid form, one tautomer usually predominates. Although a single tautomer may be described, the present invention includes all tautomers of the present compounds of formula I.

In addition, some of the compounds of the invention may form atropisomers (eg, substituted biaryls). Atropoisomers are conformational stereoisomers that occur when rotation around a single bond in a molecule is prevented or significantly slowed down by spatial interactions with other parts of the molecule, and the substituents on both ends of the single bond are asymmetrical. The interconversion of atropisomers occurs slowly enough that they can be separated and isolated under given conditions. The energy barrier to thermal racemization may be determined by the steric hindrance to the free rotation of one or more bonds forming a chiral axis.

Conventional methods for preparing/isolating individual enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate (or racemate salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC) or supercritical liquid chromatography (SLC).

Alternatively, the racemate (or racemic precursor) may be reacted with a suitable optically active compound, such as an alcohol, or, in the case where the compound contains an acidic or basic group, an acid or base such as tartaric acid or 1-phenylethylamine. The resulting diastereomeric mixture can be separated by chromatography and/or fractional crystallization, and one or both diastereoisomers converted to the corresponding pure enantiomer(s) by methods well known to one skilled in the art.

The chiral compounds of the invention (and their chiral precursors) can be obtained in enantiomerically enriched form using chromatography, typically HPLC, on an asymmetric resin with a mobile phase consisting of a hydrocarbon, typically heptane or hexane, containing from 0 to 50% isopropanol, typically , from 2 to 20% and from 0 to 5% alkylamine, usually 0.1% diethylamine. The concentration of the eluate produces a rich mixture.

Stereoisomeric conglomerates can be separated by conventional methods known to those skilled in the art; see, for example, “Stereochemistry of Organic Compounds” by E L Eliel (Wiley, New York, 1994), the description of which is incorporated herein by reference in its entirety.

The enantiomeric purity of the compounds described herein can be described in terms of enantiomeric excess (EE), which indicates the extent to which a sample contains one enantiomer in greater quantity than the other. A racemic mixture has 0% ee, while a single completely pure enantiomer has 100% ee. Similarly, diastereomeric purity can be described in terms of diastereomeric excess (di).

The present invention also includes compounds labeled with isotopes that are identical to the compounds specified in one of the presented Formulas I, but with the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature.

The isotopically labeled compounds of the invention can generally be prepared by conventional methods known to those skilled in the art, or by methods similar to those described herein, using an appropriate isotopically labeled reagent in place of the unlabeled reagent otherwise used.

Examples of isotopes that may be included in the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine such as, but not limited to, ^2H , ^3H , ^13C , ^14C , ^15N , ¹⁸ O, ¹⁷ O, ³¹ R, ³² R, ³⁵ S, ¹⁸ F and ³⁶ Cl. Certain isotopically labeled compounds of the invention, for example those incorporating radioactive isotopes such as ³ H and ¹⁴ C, can be used in drug and/or substrate tissue distribution assays. Tritium isotopes, i.e. ³ H, and carbon-14, i.e. ¹⁴ C are particularly preferred due to their ease of preparation and detection. In addition, substitution with heavier isotopes such as deuterium, e.g. ^2H may provide certain therapeutic benefits due to greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and therefore may be preferred in some circumstances. The isotopically labeled compounds of the present invention can generally be prepared by following the procedures described in the Schemes and/or in the following Examples and Preparations by replacing the non-isotope-labeled reagent with an isotope-labeled reagent.

The compounds of the invention for pharmaceutical use can be administered as crystalline or amorphous products or mixtures thereof. They can be obtained, for example, in the form of solid layers, powders or films by methods such as precipitation, crystallization, freeze-drying, spray-drying or evaporation-drying. Microwave or radiofrequency drying can be used for this purpose. The compounds of the invention may exist on a continuum of solid states ranging from completely amorphous to completely crystalline. The term "amorphous" refers to a state in which a material lacks long-range order at the molecular level and, depending on temperature, can exhibit the physical properties of a solid or a liquid. Typically, such materials do not produce distinct x-ray diffraction patterns and, although they exhibit solid-state properties, are more formally described as liquids. When heated, a transition occurs from solid to liquid properties, which is characterized by a change in state, usually of the second order (“glass transition”). The term "crystalline" refers to a solid phase in which the material has a regular, ordered internal structure at the molecular level and produces a distinct x-ray pattern with defined peaks. Such materials, when heated sufficiently, will also exhibit the properties of a liquid, but the transition from solid to liquid is characterized by a phase transition, usually first order ("melting point").

The compounds of formula I can also exist in a mesomorphic state (mesophase or liquid crystals) when exposed to suitable conditions. The mesomorphic state is intermediate between the true crystalline state and the true liquid state (melt or solution). Mesomorphism resulting from a change in temperature is described as "thermotropic" and that resulting from the addition of a second component such as water or another solvent as "lyotropic". Compounds capable of forming lyotropic mesophases are described as "amphiphilic" and consist of molecules that are ionic (such as -COO ^- Na ⁺ , -COO ^- K ⁺ or -SO ₃ ^- Na ⁺ ) or non-ionic (such as -N ^- N ⁺ (CH ₃ ) ₃ ) polar head group. For more information, see Crystals and the Polarizing Microscope by N.H. Hartshorne and A. Stuart, ^4th Edition (Edward Arnold, 1970).

Compounds of formula I may exhibit polymorphism and/or one or more types of isomerism (eg, optical, geometric or tautomeric isomerism). Compounds of formula I may also be isotopically labeled. This variation is intended for compounds of formula I, defined by their structural features and therefore included within the scope of the invention.

Therapeutic modalities and uses

The invention further relates to therapeutic methods and uses involving the administration of compounds of the invention, or pharmaceutically acceptable salts thereof, alone or in combination with other therapeutic agents or palliative agents.

In one embodiment, the invention provides a method of treating abnormal cell growth in a subject, comprising administering to the subject a therapeutically effective amount of a compound of the invention or a pharmaceutically acceptable salt thereof. In frequent embodiments, the abnormal cell growth is cancer.

In another embodiment, the invention provides a method of treating cancer in a subject, comprising administering to the subject an amount of a compound of the invention or a pharmaceutically acceptable salt thereof in combination with an amount of an additional anticancer therapeutic agent, wherein the amounts together are effective in treating said cancer.

The compounds of the invention include compounds of formula I, as described herein, or pharmaceutically acceptable salts thereof.

In yet another embodiment, the invention provides a method of inhibiting the proliferation of cancer cells in a subject, comprising administering to the subject an effective amount of a compound of the invention or a pharmaceutically acceptable salt thereof.

In another embodiment, the invention provides a method of inhibiting the invasiveness of cancer cells in a subject, comprising administering to the subject an effective amount of a compound of the invention or a pharmaceutically acceptable salt thereof.

In another embodiment, the invention relates to a method of causing the death of cancer cells in a subject, comprising administering to the subject an effective amount of a compound of the invention or a pharmaceutically acceptable salt thereof.

In yet another embodiment, the invention provides a method of enhancing vaccine therapy in a mammal, comprising administering to the mammal a therapeutically effective amount of a vaccine and further comprising administering to the mammal a therapeutically effective amount of a compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof.

In yet another embodiment, the invention provides a method of improving the ability of the immune system to eliminate a viral infection, bacterial infection or pathogen (including parasitic worms) in a subject, comprising administering to the subject an effective amount of a compound of the invention or a pharmaceutically acceptable salt thereof. The invention provides a method of enhancing immune system-mediated pathogen clearance comprising administering to a mammal a therapeutically effective amount of a compound of any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof. The method involves administering a compound of the invention as monotherapy or in combination with other agents to treat an infection or pathogen.

The compounds described herein find use in inhibiting HPK1 kinase activity. HPK1, also called mitogen-activated protein kinase kinase kinase 1 or MAP4K1, is a member of the Ste20-related series/threnonine kinase subfamily of germinal center kinases. HPK1 kinase functions as MAP4K by phosphorylating and activating MAP3K proteins, including MEKK1, MLK3 and TAK1, resulting in activation of MAPK,7nk.

HPK1 polynucleotides and polypeptides are known in the art (Hie et al. (1996) Genes Dev. 10: 2251-2264, which is incorporated herein by reference in its entirety). HPK1 polypeptides contain many conserved structural motifs. HPK1 polypeptides contain an amino-terminal Ste20-like kinase domain that spans amino acid residues 17-293, which includes an ATP-binding site from amino acid residues 23-46. The kinase domain is followed by four proline-rich (PR) motifs that serve as binding sites for SH3-containing proteins such as CrkL, Grb2, HIP-55, Gads, Nek, and Crk. The four PR motifs span amino acid residues 308–407, 394–402, 432–443, and 468–477, respectively. HPK1 becomes phosphorylated and activated in response to TCR or BCR stimulation. TCR- and BCR-induced tyrosine phosphorylation at position 381, located between PR1 and PR2, mediates binding to SLP-76 in T cells or BLNK in B cells through the SLP-76 or BLNK SH2 domain, and is required for kinase activation. The citron homology domain found at the C terminus of HPK1, approximately spanning residues 495–800, may act as a regulatory domain and may be involved in macromolecular interactions.

The compounds described herein bind directly to HPK1 and inhibit its kinase activity. In some embodiments, compounds described herein reduce, inhibit, or otherwise reduce HPK1-mediated phosphorylation of SLP76 and/or Gads. The compounds described herein may or may not be a specific inhibitor of HPK1. A specific HPK1 inhibitor reduces the biological activity of HPK1 by an amount that is statistically greater than the inhibitory effect of the inhibitor on any other protein (eg, other series/threonine kinases). In some embodiments, the compounds described herein specifically inhibit the serine/threonine kinase activity of HPK1.

The compounds described herein can be used in a method of inhibiting HPK1. Such methods involve contacting HPK1 with an effective amount of a compound described herein. The term “contact” means bringing the compound into sufficiently close contact with an isolated HPK1 enzyme or a cell expressing HPK1 (eg, T cell, B cell, dendritic cell) such that the compound is able to bind HPK1 and inhibit its activity. The compound can contact HPK1 in vitro or in vivo by administering the compound to a subject.

Any method known in the art for measuring HPK1 kinase activity can be used to determine whether HPK1 has been inhibited, including in vitro kinase assays, immunoblots with antibodies specific for phosphorylated targets of HPK1, such as SLP76 and Gads, or measurement subsequent biological effect of HPK1 kinase activity, such as recruitment of 14-3-3 proteins to phosphorylated SLP7 and Gads, release of the SLP76-Gads-14-3-3 complex from LAT-containing microclusters, or activation of T or B cells.

The compounds described herein can be used to treat an HPK1l-dependent disorder. As used herein, the term “HPK1-dependent disorder” is a pathological condition in which HPK1 activity is required for the initiation or maintenance of the pathological condition.

The compounds described herein also find use for enhancing the immune response in a subject in need thereof. Such methods involve administering an effective amount of a presently described compound (ie, any of the compounds of Formula I or a pharmaceutically acceptable salt, prodrug, metabolite or derivative thereof). The term "enhancement of the immune response" refers to the improvement of any immunogenic response to an antigen. Non-limiting examples of improving the immunogenic response to an antigen include enhanced dendritic cell maturation or migration, enhanced T cell activation (e.g. CD4 T cells, CD8 T cells), T cell enhancement (e.g. CD4 T cells, CD8 T cells ). T cell proliferation, enhanced B cell proliferation, increased T cell and/or B cell survival, improved antigen presentation by antigen presenting cells (e.g., dendritic cells), improved antigen clearance, increased production of cytokines by T cells (e.g., interleukin- 2), increased resistance to immunosuppression induced by prostaglandin E2 or adenosine, and increased priming and/or cytolytic activity of CD8 T cells. In some embodiments, CD8 T cells in a subject have increased priming, activation, proliferation, and/or cytolytic activity compared to prior administration of a compound of Formula I or a pharmaceutically acceptable salt, prodrug, metabolite, or derivative thereof. In some embodiments, CD8 T cell priming is characterized by increased expression of CD44 and/or increased cytolytic activity in CD8 T cells. In some embodiments, CD8 T cell activation is characterized by an increased frequency of IFNy ⁺ CD8 T cells. In some embodiments, the CD8 T cell is an antigen-specific T cell.

In some embodiments, antigen presenting cells in a subject have increased maturation and activation compared to before administration of a compound of Formula I or a pharmaceutically acceptable salt, prodrug, metabolite, or derivative thereof. In some embodiments, the antigen presenting cells are dendritic cells. In some embodiments, maturation of antigen presenting cells is characterized by an increased frequency of CD83 ⁺ dendritic cells. In some embodiments, activation of antigen presenting cells is characterized by increased expression of CD80 and CD86 on dendritic cells.

TCR engagement leads to the activation of HPK1, which functions as a negative regulator of the TCR-induced AP-1 response pathway. HPK1 is thought to negatively regulate T cell activation by reducing the persistence of signaling microclusters by phosphorylating SLP76 at Ser376 (Di Bartolo et al. (2007), 7EM 204:681-691) and Gads at Thr254, leading to the recruitment of 14-3- 3, which bind to phosphorylated SLP76 and Gads, releasing the SLP76-Gads-14-3-3 complex from LAT-containing microclusters, leading to T cell dysfunction, including anergy and exhaustion (Lasserre et al. (2011).), 7 Cell Biol 195(5): 839-853). The term "dysfunction" in the context of immune dysfunction refers to a state of reduced immune response to antigenic stimulation. The term includes common elements of both wasting and/or anergy, in which antigen recognition can occur but the subsequent immune response is ineffective in controlling infection or tumor growth.

The term “dysfunctional” as used herein also includes refractoriness or unresponsiveness to antigen recognition, particularly, impaired ability to translate antigen recognition into subsequent T cell effector functions such as proliferation, cytokine production (eg, IL-2, gamma IFN) and/or destruction of target cells.

The term "anergy" refers to a state of unresponsiveness to antigenic stimulation resulting from incomplete or insufficient signals delivered through the T-cell receptor (eg, increased intracellular Ca in the absence of ras activation). T cell anergy can also occur when stimulated by antigen in the absence of costimulation, causing the cell to become refractory to subsequent activation by antigen even in the context of costimulation. The unresponsive state can often be overcome by the presence of interleukin-2. Anergic T cells do not undergo clonal expansion and/or acquire effector functions.

The term "exhaustion" refers to T cell exhaustion as a state of T cell dysfunction that occurs due to persistent TCR signaling that occurs during many chronic infections and cancer. It differs from anergy in that it does not arise from incomplete or insufficient signaling, but from sustained signaling. It is defined by poor effector function, persistent expression of inhibitory receptors, and a transcriptional state distinct from that of functional effector or memory T cells. Depletion prevents optimal control of infection and tumors. Exhaustion can be the result of both external negative regulatory pathways (for example, immunoregulatory cytokines) and internal negative regulatory (co-stimulatory) pathways of the cell (PD-1, B7-H3, B7-H4, etc.).

“Enhance T cell function” means inducing, causing, or stimulating a T cell to have sustained or enhanced biological function, or renewing or reactivating exhausted or inactive T cells. Examples of enhanced T cell function include: increased secretion of cytokines (eg, interferon gamma, IL-2, IL-12, and TNF-alpha), increased proliferation, increased sensitivity to antigens (eg, clearance of viruses, pathogens, or tumors) versus with such preintervention levels, and increased production of effector granules by CD8 T cells such as granzyme B.

Accordingly, the compounds of Formula I described herein, or pharmaceutically acceptable salts, prodrugs, metabolites or derivatives thereof, are useful for the treatment of disorders associated with T cell dysfunction. "T cell dysfunctional disorder" is a disorder or condition of T cells characterized by a reduced response to antigenic stimulation. In a specific embodiment, the T cell dysfunctional disorder is a disorder that is specifically associated with increased HPK1 kinase activity. In another embodiment, a dysfunctional T cell disorder is a disorder in which the T cells are energetic or have a reduced ability to secrete cytokines, proliferate, or perform cytolytic activity. In a particular aspect, reduced reactivity results in ineffective control of the pathogen or tumor expressing the immunogen. Examples of T cell dysfunctional disorders characterized by T cell dysfunction include unresolved acute infection, chronic infection, and tumor immunity.

Thus, the compounds described herein can be used to treat conditions in which increased immunogenicity is desired, for example, to enhance tumor immunogenicity for the treatment of cancer. "Immunogenicity" refers to the ability of a particular substance to elicit an immune response. Tumors are immunogenic, and increasing tumor immunogenicity promotes clearance of tumor cells through the immune response.

"Tumor immunity" refers to the process by which tumors evade immune recognition and clearance. Thus, as a therapeutic concept, tumor immunity is “treated” when such avoidance is weakened and tumors are recognized and attacked by the immune system. Examples of tumor recognition include tumor binding, tumor shrinkage, and tumor removal.

In one aspect, provided herein is a method of treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound of Formula I or a pharmaceutically acceptable salt, prodrug, metabolite, or derivative thereof. In some embodiments, the subject has melanoma. Melanoma can be early or late stage. In some embodiments, the subject has colorectal cancer. Colorectal cancer can be early or late stage. In some embodiments, the subject has non-small cell lung cancer. Non-small cell lung cancer can be early or late stage. In some embodiments, the subject has pancreatic cancer. Pancreatic cancer can be early or late stage. In some embodiments, the subject has a hematologic malignancy.

Hematologic malignancy can be early or late stage. In some embodiments, the subject has ovarian cancer. Ovarian cancer can be early or late stage. In some embodiments, the subject has breast cancer. Breast cancer can be early stage or advanced stage. In some embodiments, the subject has renal cell carcinoma. Renal cell carcinoma can be early stage or advanced stage. In some embodiments, the cancer is characterized by increased levels of T cell infiltration.

In some embodiments, the treatment results in a sustained response in the subject after treatment is discontinued. "Sustained response" refers to the sustained effect of reducing tumor growth after stopping treatment. For example, the tumor size may remain the same or smaller than the size at the beginning of the administration phase.

The treatments described herein may result in a partial or complete response. As used herein, the term “complete response” or “CR” refers to the disappearance of all target lesions; “partial response” or “PR” refers to at least a 30% reduction in the sum of the longest diameters (SLD) of the target lesions, based on the initial SLD; and “stable disease” or “SD” refers to neither a sufficient reduction in target lesions to qualify as PR nor a sufficient increase to qualify as PD, taking as the reference the lowest SLD since the start of treatment. As used herein, the term “overall response rate” (ORR) refers to the sum of the complete response rate (CR) and the partial response rate (PR).

The treatments described herein may result in an increase in progression-free survival and overall survival of a subject treated with an HPK1 antagonist. As used herein, the term “progression-free survival” (PFS) refers to the length of time during and after treatment during which the disease being treated (eg, cancer) does not worsen. Progression-free survival may include the amount of time patients have a complete or partial response, as well as the amount of time patients have stable disease.

As used herein, the term "overall survival" refers to the proportion of subjects in a group who are likely to be alive after a specified period of time.

In some embodiments of the methods provided herein, the abnormal cell growth is a cancer, where the cancer is selected from the group consisting of breast cancer, ovarian cancer, bladder cancer, uterine cancer, prostate cancer, lung cancer (including NSCLC, SCLC, squamous cell carcinoma or adenocarcinoma), esophageal cancer, head and neck cancer, colorectal cancer, kidney cancer (including RCC), liver cancer (including HCC), pancreatic cancer, stomach cancer (eg, gastric) and thyroid cancer. In additional embodiments of the methods provided herein, the cancer is selected from the group consisting of breast cancer, ovarian cancer, bladder cancer, uterine cancer, prostate cancer, lung cancer, esophageal cancer, liver cancer, pancreatic cancer, and cancer stomach.

In some embodiments, the cancer is selected from the group consisting of breast cancer and ovarian cancer.

In some embodiments, the cancer is ovarian cancer.

In other embodiments, the cancer is breast cancer, including, for example, EI-positive/NI-positive breast cancer, HERP2-negative breast cancer; ER-positive/HR-positive breast cancer, HEI2-positive breast cancer; triple negative breast cancer (TNBC); or inflammatory breast cancer. In some embodiments, the breast cancer is endocrine-resistant breast cancer, trastuzumab-resistant breast cancer, or breast cancer demonstrating primary or acquired resistance to CDK4/CDK6 inhibition. In some embodiments, the breast cancer is advanced or metastatic breast cancer.

In some embodiments, a compound of the invention is administered as first-line therapy. In other embodiments, a compound of the invention is administered as second (or later) line therapy. In some embodiments, a compound of the invention is administered as second (or later) line therapy following treatment with an endocrine therapeutic agent and/or a CDK4/CDK6 inhibitor. In some embodiments, a compound of the invention is administered as second (or later) line therapy following treatment with an endocrine therapeutic agent. In some embodiments, a compound of the invention is administered as second (or later) line therapy following treatment with a CDK4/CDK6 inhibitor. In some embodiments, a compound of the invention is administered as second (or later) line therapy after treatment with one or more chemotherapy regimens, for example, including taxanes or platinum agents. In some embodiments, a compound of the invention is administered as second (or later) line therapy following treatment with HER2-targeting agents, such as trastuzumab.

The terms "abnormal cell growth" and "hyperproliferative disorder" are used interchangeably in this application.

“Abnormal cell growth” as used herein, unless otherwise noted, refers to cell growth that is independent of normal regulatory mechanisms (eg, loss of contact inhibition). Abnormal cell growth may be benign (non-cancerous) or malignant (cancerous).

Abnormal cell growth includes abnormal growth of tumors resistant to endocrine therapy, HER2 antagonists, or CDK4/6 inhibition.

The term "additional anticancer therapeutic agent" as used herein means any one or more therapeutic agents, other than a compound of the invention, that are or may be used to treat cancer, such as agents derived from the following classes: mitotic inhibitors, alkylating agents , antimetabolites, antitumor antibiotics, topoisomerase I and II inhibitors, plant alkaloids, hormonal agents and antagonists, growth factor inhibitors, radiation, protein tyrosine kinase and/or series/threonine kinase inhibitors, cell cycle inhibitors, biological response modifiers, enzyme inhibitors, antisense oligonucleotides or derivatives oligonucleotides, cytotoxic and immuno-oncological agents (immuno-oncological agents include monoclonal antibodies, bispecific antibodies, cytokines, CAR-t cells).

As used herein, the term “cancer” refers to any malignant and/or invasive growth or tumor caused by abnormal cell growth. Cancer includes solid tumors, named for the type of cells that form them, cancers of the blood, bone marrow, or lymphatic system. Examples of solid tumors include sarcomas and carcinomas. Blood cancers include, but are not limited to, leukemia, lymphoma and myeloma. Cancer also includes primary cancer, which occurs in a specific location in the body, metastatic cancer, which has spread from the place where it started to other parts of the body, recurrence of the original primary cancer after remission, and second primary cancer, which is a new primary cancer in a person who has a history of another type of cancer.

In some embodiments of the methods provided herein, the cancer is selected from the group consisting of breast cancer, ovarian cancer, bladder cancer, uterine cancer, prostate cancer, lung cancer, esophageal cancer, liver cancer, pancreatic cancer, and cancer stomach.

Dosage forms and schemes

Administration of the compounds of the invention can be accomplished by any method that offers delivery of the compounds to the site of action. These routes include oral route of administration, intraduodenal route of administration, parenteral injection (including intravenous, subcutaneous, intramuscular, intravascular or infusion), topical and rectal administration.

Dosing regimens may be adjusted to provide the optimal desired response. For example, a single bolus may be administered, multiple divided doses may be administered over time, or the dose may be proportionally decreased or increased as required by the therapeutic situation. It is particularly advantageous to formulate parenteral compositions in unit dosage form for ease of administration and uniformity of dosage. Unit dosage form as used herein refers to physically discrete units suitable as unit dosages for the mammal to be treated; each unit contains a predetermined amount of active compound calculated to produce the desired therapeutic effect, in combination with the required pharmaceutical carrier. The specification of the unit dosage forms of the invention is dictated by and directly depends on (a) the unique characteristics of the chemotherapeutic agent and the particular therapeutic or prophylactic effect to be achieved, and (b) the limitations inherent in the art of formulating such an active compound for the treatment of sensitivity in individuals.

Thus, one skilled in the art will appreciate, based on the disclosure herein, that the dosage and dosage schedule will be adjusted in accordance with methods well known in the art of therapy. That is, the maximum tolerated dose can be easily determined, and the effective amount to provide a detectable therapeutic effect in a patient, as well as the time requirements for administering each agent to provide a detectable therapeutic effect in a patient, can be determined. Accordingly, while examples of specific dosages and dosage schedules are provided herein, these examples do not in any way limit the dosages and dosage schedules that may be provided to a patient in the practice of the present invention.

It should be noted that dosage amounts may vary depending on the type and severity of the condition being alleviated and may include single or multiple doses. In addition, it should be understood that for any particular subject, specific dosage regimens should be adjusted over time in accordance with the individual needs and professional judgment of the person administering or supervising the administration of the compositions, and that the dosage ranges set forth herein are only approximate and are not intended to limit the scope or application of the claimed composition. For example, dosages may be adjusted based on pharmacokinetic or pharmacodynamic parameters, which may include clinical effects such as toxic effects and/or laboratory parameters. Thus, the present invention covers intra-patient dose escalation as determined by one skilled in the art. Determination of suitable dosages and schedules of administration of a chemotherapeutic agent is well known in the relevant art, and one skilled in the art will understand that they are covered as soon as the ideas described herein are proposed.

The amount of a compound of the invention administered will depend on the subject being treated, the severity of the disorder or condition, the rate of administration, the location of the compound, and the discretion of the attending physician. However, an effective dose is in the range of from about 0.001 to about 100 mg per kg of body weight per day, preferably from about 1 to about 35 mg/kg/day, as a single or divided dose. For a 70 kg person this would be from about 0.05 to about 7 g/day, preferably from about 0.1 to about 2.5 g/day. In some cases, dose levels below the lower end of the above range may be more than sufficient, while in other cases even larger doses can be used without causing any harmful side effects, provided such larger doses are first divided into several doses. small doses to be administered throughout the day.

Compositions and routes of administration

As used herein, the term “pharmaceutically acceptable carrier” refers to a carrier or diluent that does not cause significant irritation to the body and does not interfere with the biological activity and properties of the compound administered.

The pharmaceutically acceptable carrier may include any conventional pharmaceutical carrier or excipient. The choice of carrier and/or excipient will depend largely on factors such as the particular route of administration, the effect of the carrier or excipient on solubility and stability, and the nature of the dosage form.

Suitable pharmaceutical carriers include inert diluents or excipients, water and various organic solvents (such as hydrates and solvates). Pharmaceutical compositions may, if desired, contain additional ingredients such as flavoring agents, binders, excipients and the like. Thus, for oral administration, tablets containing various excipients such as citric acid can be used together with various disintegrants such as starch, alginic acid and some complex silicates, and with binding agents such as sucrose, gelatin and acacia. Examples of excipients include, without limitation, calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols. In addition, lubricants such as magnesium stearate, sodium lauryl sulfate and talc are often used for tableting. Solid compositions of a similar type can also be used in soft and hard filled gelatin capsules. Thus, non-limiting examples of materials include lactose or milk sugar and high molecular weight polyethylene glycols. When aqueous suspensions or elixirs are desired for oral administration, the active compound contained therein may be combined with various sweeteners or flavoring agents, coloring agents or pigments and, if desired, emulsifiers or suspending agents together with diluents such as water, ethanol, propylene glycol, glycerin or their combinations.

The pharmaceutical composition may be, for example, in a form suitable for oral administration, as a tablet, capsule, pill, powder, sustained release formulation, suspension in solution, for parenteral injection as a sterile solution, suspension or emulsion, for topical administration to in the form of an ointment or cream, or for rectal administration in the form of suppositories.

Typical forms for parenteral administration include solutions or suspensions of the active compounds in sterile aqueous solutions, for example, aqueous solutions of propylene glycol or dextrose. If desired, such dosage forms may be suitably buffered.

The pharmaceutical composition may be in unit dosage forms suitable for single administration in precise dosages.

Pharmaceutical compositions suitable for delivering the compounds of the invention and methods for their preparation will be apparent to those skilled in the art. Such compositions and methods for preparing them can be found, for example, in Remington's Pharmaceutical Sciences, ^19th Edition (Mack Publishing Company, 1995), the disclosure of which is incorporated herein by reference in its entirety.

The compounds of the invention can be administered orally. Oral administration may involve ingestion, so that the compound enters the gastrointestinal tract, or buccal or sublingual administration may be used, whereby the compound enters the bloodstream directly from the mouth.

Formulations suitable for oral administration include solid formulations such as tablets, capsules containing particles, liquids or powders, lozenges (including liquid-filled), chewable tablets, multi- and nanoparticles, gels, solid solution, liposomes, films (including mucoadhesive ), capsules, sprays and liquid formulations.

Liquid formulations include suspensions, solutions, syrups and elixirs. Such formulations may be used as fillers in soft or hard capsules, and typically include a carrier, for example water, ethanol, polyethylene glycol, propylene glycol, methylcellulose or a suitable oil, and one or more emulsifiers and/or suspending agents. Liquid formulations can also be prepared by reconstituting a solid, for example from a sachet.

The compounds of the invention may also be used in rapidly dissolving and rapidly disintegrating dosage forms, such as those described in Expert Opinion in Therapeutic Patents, 11(6), 981-986 Liang and Chen (2001), the disclosure of which is incorporated herein by reference in its entirety .

For tablet dosage forms, depending on the dose, the drug can be from 1% by weight. up to 80% wt. dosage form, more typically from 5% wt. up to 60% wt. dosage form. In addition to the drug, the tablets usually contain a disintegrant. Examples of disintegrants include sodium starch glycolate, sodium carboxymethylcellulose, calcium carboxymethylcellulose, croscarmellose sodium, crospovidone, polyvinylpyrrolidone, methylcellulose, microcrystalline cellulose, lower alkyl-substituted hydroxypropylcellulose, starch, pregelatinized starch and sodium alginate. As a rule, the baking powder is from 1% of the mass. up to 25% wt. preferably from 5% wt. up to 20% wt. dosage form.

Binding agents are commonly used to impart cohesive properties to tablet formulations. Suitable binding agents include microcrystalline cellulose, gelatin, sugars, polyethylene glycol, natural and synthetic gums, polyvinylpyrrolidone, pregelatinized starch, hydroxypropylcellulose and hydroxypropylmethylcellulose. The tablets may also contain diluents such as lactose (monohydrate, spray-dried monohydrate, anhydrous and the like), mannitol, xylitol, dextrose, sucrose, sorbitol, microcrystalline cellulose, starch and dibasic calcium phosphate dihydrate.

Tablets may also optionally include surfactants such as sodium lauryl sulfate and polysorbate 80, and glidants such as silicon dioxide and talc. Surface-active agents, if present, are typically present in amounts ranging from 0.2% by weight. up to 5% wt. tablets and glidants usually range from 0.2% wt. up to 1% wt. pills.

Tablets also typically contain lubricating agents such as magnesium stearate, calcium stearate, zinc stearate, sodium stearyl fumarate, and mixtures of magnesium stearate and sodium lauryl sulfate. Lubricants are typically present in amounts ranging from 0.25% by weight. up to 10% wt. preferably from 0.5% wt. up to 3% wt. pills.

Other traditional ingredients include antioxidants, colors, flavors, preservatives, and flavor-masking agents.

Examples of tablets contain up to about 80 wt.%. drug, from about 10% of the mass. up to about 90% of the mass. binding agent, from about 0% wt. up to about 85% wt. diluent, from about 2% by weight. up to about 10% wt. baking powder and from about 0.25 wt.%. up to about 10% by weight, lubricant.

Tablet mixtures can be compressed directly or using a roller to form tablets. Tablet mixtures or portions of mixtures may alternatively be prepared by wet, dry or melt granulation, melt-frozen or extruded prior to tableting. The final composition may include one or more layers, and may be coated or uncoated; or encapsulated.

The composition of the tablets is discussed in detail in "Pharmaceutical Dosage Forms: Tablets, Vol. 1", H. Lieberman and L. Lachman, Marcel Dekker, NY, NY, 1980 (1SBN 0-8247-6918-X), the description of which is incorporated herein in its entirety description by reference.

Solid formulations for oral administration can be formulated for immediate and/or modified release. Modified release formulations include delayed, extended, pulsed, controlled, targeted and programmed release.

Suitable modified release formulations are described in US Pat. No. 6,106,864. Details of other suitable release technologies such as high energy dispersions and osmotic and coated particles can be found in Verma et al., Pharmaceutical Technology On-line, 25(2), 1-14 (2001). The use of chewing gum to achieve controlled release is described in WO 00/35298. The descriptions of these references are incorporated herein by reference in their entirety.

Parenteral administration

The compounds of the invention can also be administered directly into the bloodstream, into a muscle or into an internal organ. Suitable means for parenteral administration include intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, intrasternal, intracranial, intramuscular and subcutaneous administration. Suitable devices for parenteral administration include needle (including microneedle) injectors, needleless injectors, and infusion methods.

Formulations for parenteral administration are typically aqueous solutions that may contain excipients such as salts, carbohydrates and buffering agents (preferably pH 3 to 9), but for some applications they may be more suitably formulated as a sterile, non-aqueous solution or in dried form for use in combination with a suitable carrier such as sterile pyrogen-free water.

The preparation of parenteral formulations under sterile conditions, for example by lyophilization, can be easily accomplished using standard pharmaceutical techniques well known to those skilled in the art.

The solubility of the compounds of the invention used in the preparation of parenteral solutions can be increased by the use of suitable formulation techniques, such as the addition of solubility enhancing agents.

Formulations for parenteral administration can be formulated for immediate and/or modified release. Modified release formulations include delayed, extended, pulsed, controlled, targeted and programmed release. Thus, the compounds of the invention may be formulated as a solid, semi-solid or thixotropic liquid for administration as an implanted depot providing modified release of the active compound. Examples of such formulations include drug-eluting stents and PGLA microspheres.

The compounds of the invention can also be administered topically to the skin or mucous membrane, that is, dermally or transdermally. Typical formulations for this purpose include gels, hydrogels, lotions, solutions, creams, ointments, powders, dressings, foams, films, skin patches, wafers, implants, sponges, fibers, bandages and microemulsions. Liposomes can also be used. Typical carriers include alcohol, water, mineral oil, liquid petrolatum, white petrolatum, glycerin, polyethylene glycol and propylene glycol. Penetration enhancers may be included; see, for example, 7 Pharm Sci, 88 (10), 955-958 by Finnin and Morgan (October 1999). Other methods of local administration include delivery by electroporation, iontophoresis, phonophoresis, sonophoresis, and microneedle or non-needle injection (eg, Powderject™, Bioject™, etc.). The descriptions of these references are incorporated herein by reference in their entirety.

Formulations for topical administration can be formulated for immediate and/or modified release. Modified-release drugs include delayed, extended-release, pulsed, controlled, targeted, and programmed release.

The compounds of the invention can also be administered intranasally or by inhalation, typically in the form of a dry powder (either alone or as a mixture, for example in a dry mixture with lactose, or as particulates of a mixed component, for example in a mixture with phospholipids such as phosphatidylcholine) from a dry powder inhaler or as an aerosol spray from a pressurized can, pump, spray, nebulizer (preferably an atomizer using electrohydrodynamics to produce a fine mist) or nebulizer, with or without the use of a suitable propellant such as 1,1 ,1,2-tetrafluoroethane or 1,1,1,2,3,3,3-heptafluoropropane. For intranasal use, the powder may include a bioadhesive such as chitosan or cyclodextrin.

The pressurized container, pump, spray, nebulizer or nebulizer contains a solution or suspension of the compound(s) of the invention containing, for example, ethanol, an aqueous solution of ethanol or a suitable alternative agent for dispersing, solubilizing or prolonging the release of the active substance, propellant(s) in as a solvent and an optional surfactant such as sorbitan trioleate, oleic acid or oligolactic acid.

Before use as a dry powder or suspension, the drug product is micronized to a size suitable for delivery by inhalation (usually less than 5 microns). This can be achieved by any suitable comminution method, such as spiral jet milling, fluidized bed jet milling, supercritical fluid nanoparticle treatment, high pressure homogenization, or spray drying.

Capsules (made, for example, from gelatin or HPMC), blisters and cartridges for use in an inhaler or insufflator can be formulated to contain a powder mixture of a compound of the invention, a suitable powder base such as lactose or starch, and a performance modifier, such as leucine, mannitol or magnesium stearate. Lactose may be anhydrous or in monohydrate form, preferably the latter form. Other suitable excipients include dextran, glucose, maltose, sorbitol, xylitol, fructose, sucrose and trehalose.

A suitable solution formulation for use in a nebulizer using electrohydrodynamics to produce a fine mist may contain from 1 μg to 20 mg of a compound of the invention per stroke, and the working volume may vary from 1 μl to 100 μl. A typical formulation includes a compound of the invention, propylene glycol, sterile water, ethanol and sodium chloride. Alternative solvents that can be used instead of propylene glycol include glycerin and polyethylene glycol.

Suitable flavoring agents, such as menthol and levomenthol, or sweeteners, such as saccharin or sodium saccharin, can be added to those compositions of the invention that are intended for inhalation/intranasal administration.

Formulations for inhalation/intranasal administration can be formulated to provide immediate and/or modified release using, for example, poly(DL-lactic-coglycolic acid (PGLA). Modified release compositions include delayed, sustained release, pulsed, controlled, targeted and programmed release.

In the case of dry powder and aerosol inhalers, the dosage unit is determined by a valve that delivers a metered amount. Units of the invention are typically designed to deliver a metered dose or "spray" containing the desired amount of a compound of the invention. The total daily dose may be administered as a single dose or, more commonly, in divided doses throughout the day.

The compounds of the invention can be administered rectally or vaginally, for example, in the form of a suppository, pessary or enema. Cocoa butter is a traditional suppository base, but various alternatives can be used if necessary.

Formulations for rectal/vaginal administration may be formulated for immediate and/or modified release. Modified release formulations include delayed, extended, pulsed, controlled, targeted and programmed release.

The compounds of the invention can also be administered directly into the eye or ear, typically in the form of drops of a micronized suspension or solution in isotonic, pH-adjusted sterile saline. Other formulations suitable for ocular and auricular administration include ointments, biodegradable (eg, absorbable gel sponges, collagen) and non-biodegradable (eg, silicone) implants, wafers, lenses, and particle or vesicle systems such as niosomes or liposomes. A polymer such as cross-linked polyacrylic acid, polyvinyl alcohol, hyaluronic acid, a cellulosic polymer such as hydroxypropyl methylcellulose, hydroxyethylcellulose or methylcellulose, or a heteropolysaccharide polymer such as gellan gum may be included along with a preservative such as benzalkonium chloride. Such formulations can also be delivered using iontophoresis.

Formulations for ocular/ear administration may be formulated for immediate and/or modified release. Modified-release drugs include delayed, sustained, pulsed, controlled, targeted, or programmed release.

Other technologies

The compounds of the invention can be combined with soluble macromolecular compounds, such as cyclodextrin and suitable derivatives thereof, or polyethylene glycol containing polymers, to improve their solubility, dissolution rate, taste masking, bioavailability and/or stability for use in any of the above dosing regimens.

Drug-cyclodextrin complexes are generally suitable for most dosage forms and routes of administration. Both inclusion and non-inclusion complexes can be used. As an alternative to direct complexation with the drug, cyclodextrin can be used as an auxiliary additive, i.e. as a carrier, diluent or solubilizer. The most commonly used for these purposes are alpha, beta and gamma cyclodextrins, examples of which can be found in PCT Publication Nos. WO 91/11172, WO 94/02518 and WO 98/55148, the description of which is incorporated herein by reference in its entirety.

Dosage

The amount of active compound administered will depend on the subject being treated, the severity of the disorder or condition, the rate of administration, the location of the compound, and the discretion of the attending physician. However, an effective dose will generally range from about 0.001 to about 100 mg per kg of body weight per day, preferably from about 1 to about 35 mg/kg/day, as a single or divided dose. For a 70 kg person this would be from about 0.05 to about 7000 mg/day, preferably from about 0.1 to about 2500 mg/day. In some cases, dose levels below the lower end of the above range may be more than sufficient, while in other cases even larger doses can be used without causing any harmful side effects, with such large doses usually divided into several smaller doses for administration throughout the day.

Sets

Since it may be desirable to administer a combination of active compounds, for example, for the purpose of treating a particular disease or condition, the present invention includes two or more pharmaceutical compositions, at least one of which contains a compound according to the invention, which can be conveniently combined into in the form of a kit suitable for the joint administration of compositions. Thus, a kit of the invention includes two or more separate pharmaceutical compositions, at least one of which contains a compound of the invention, and means for separately storing said compositions, such as a container, a divided bottle or a divided foil pouch. An example of such a set is the known blister pack used for packaging tablets, capsules and the like.

The kit of the invention is particularly suitable for administering different dosage forms, for example oral and parenteral, for administering individual compositions at different dosing intervals, or for titrating individual compositions against each other. To facilitate compliance, the kit usually includes instructions for use and may be equipped with a memory device.

Combination therapy

As used herein, the term “combination therapy” refers to the administration of a compound of the invention together with at least one additional pharmaceutical or drug agent (eg, an anticancer agent, a vaccine, an antibacterial agent, an antiviral agent, or an antiparasitic agent), either sequentially or simultaneously.

As noted above, the compounds of the invention can be used in combination with one or more additional agents, such as anticancer agents. The effectiveness of the compounds of the invention in some tumors may be enhanced by combination with other approved or investigational anticancer therapies, for example, radiation therapy, surgery, chemotherapeutic agents, targeted therapies, agents that inhibit other signaling pathways that are dysregulated in tumors, and other agents. immune enhancing agents such as PD-1 antagonists and the like.

When combination therapy is used, one or more additional agents can be administered sequentially or simultaneously with a compound of the invention. In one embodiment, the additional agent is administered to the mammal (eg, human) prior to administration of the compound of the invention. In another embodiment, the additional agent is administered to a mammal (eg, human) after administration of a compound of the invention. In another embodiment, the additional agent is administered to a mammal (eg, a human) simultaneously with the administration of a compound of the invention.

The invention also relates to a pharmaceutical composition for the treatment of abnormal cell growth in a mammal, including a human, which contains an amount of a compound of the invention as defined above (including hydrates, solvates, polymorphs, isomers, prodrugs and/or metabolites of the said compound or pharmaceutically acceptable salts thereof) in combination with one or more (preferably one to three) anticancer therapeutic agents.

In specific embodiments, a compound of the invention may be administered in combination with one or more: targeted agents, such as PI3, mTOR, PARP, Kras, IDO, TDO, ALK, ROS, MEK, VEGF, FLT3, AXL, R0R2, EGFR, FGFR, Src/Al, RTK/Ras, Myc, Raf, PDGF, AKT, c-Kit, erbB, CDK2, CDK4, CDK4/CDK6, CDK5, CDK7, CDK9, SMO, CXCR4, HER2, GLS1, EZH2 or Hsp90, or immunomodulatory agents such as PD-1 antagonists, PD-L1 antagonists, CTLA-4 antagonists, OX40 agonists, 4-1 BB agonists or CD80 agonists.

In other embodiments, a compound of the present invention may be administered in combination with a standard of care such as tamoxifen, docetaxel, paclitaxel, cisplatin, capecitabine, gemcitabine, vinorelbine, exemestane, letrozole, fulvestrant, anastrozole, or trastuzumab.

Synthesis methods

The compounds of formula I can be prepared by the methods described below, together with synthetic methods known in the field of organic chemistry, or modifications and transformations known to those skilled in the art. The starting materials used herein are commercially available or can be prepared by conventional methods known in the art [such as those described in standard references such as Compendium of Organic Synthetic Methods, Vol.I-XIII (published by Wi1ey -Interscience)]. Preferred methods include, but are not limited to, the methods described below.

During any of the following synthetic sequences, it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules in question. This can be achieved using conventional protecting groups, such as those described in T. W. Greene, Protective Groups in Organic Chemistry, 7ohn Wiley & Sons, 1981; T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Chemistry, 7ohn Wiley & Sons, 1991; and T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Chemistry, 7ohn Wiley & Sons, 1999, which are incorporated herein by reference.

The compounds of Formula I or pharmaceutically acceptable salts thereof can be prepared in accordance with the reaction schemes discussed herein below. Unless otherwise noted, substituents in the diagrams have the meanings given above.

Isolation and purification of the products is carried out using standard methods known to those skilled in the field of chemistry.

One skilled in the art will appreciate that the various symbols, superscripts, and subscripts used in the diagrams, methods, and examples are for convenience of presentation and/or to reflect the order in which they are presented in the diagrams and are not intended to necessarily correspond to the symbols. , superscripts or subscripts in the attached claims. Moreover, one skilled in the art will appreciate that in many cases these compounds are mixtures and enantiomers that can be separated at various stages of the synthesis schemes using conventional methods such as, but not limited to, crystallization, normal phase chromatography , reverse phase chromatography and chiral chromatography, to obtain individual enantiomers. The schemes represent methods used in the synthesis of the compounds of the present invention. They do not limit the scope of the invention in any way.

The compounds of the invention are prepared in accordance with the standard procedures described herein and modifications thereof known to those skilled in the art.

The following abbreviations are used in the examples: "Ac" means acetyl, "ACN" means acetonitrile, "BOC", "Boc" or "boc" means N-tert-butoxycarbonyl, "Bu" means butyl, "tBu" means tert-butyl, "DCM" (CH ₂ Cl ₂ ) means methylene chloride, "di" means diastereomeric excess, "DMF" means N,N-dimethylformamide, "DMSO" means dimethyl sulfoxide, "ee" means enantiomeric excess, "Et" means ethyl, "EtOAc"" means ethyl acetate, "EtOH" means ethanol, "i-Pr" or " ⁱ Pr" means isopropyl, "Me" means methyl, "MeOH" means methanol, "MS" means mass spectrometry, "MTBE" means methyl tert -butyl ether, "Ph" means phenyl, "THF" means tetrahydrofuran, "SLC" means supercritical liquid chromatography, "TLC" means thin layer chromatography, "Rf" means retention factor, "~" means approximately, "RT" means room temperature , which includes the ambient temperature (usually 20°C to 25°C), "h" means hours, "min" means minutes, "eq" means equivalents, "sat." means saturated.

The compounds of formula I can be prepared by the methods described in the general methods presented below, or by conventional modifications thereof. Unless otherwise indicated, substituents in the methods are as defined herein. Although R ⁴ is found in compounds of formula I, it will be clear from the intermediates used in the appropriate synthetic route whether R ⁴ is R ⁴ -i or R ⁴ -ii. The present invention also covers any one or more of these methods for preparing the compounds of Formula I, in addition to any new intermediates used therein. One skilled in the art will appreciate that the following reactions can be cooled, thermally heated, microwave heated, or carried out under flow chemistry conditions. In addition, it will be understood that it may be necessary or desirable to carry out the transformations in an order different from the order described in the methods, or to modify one or more transformations to obtain the desired compound of the invention.

Method A

Method A is the preparation of compounds of formula I wherein R ^3b is hydrogen. In Method A, cross-coupling or nucleophilic aromatic substitution between the desired coupling partner or amine, provided that R ¹ is as defined in Formula I (or a protected variant thereof), gives A-2. For dichloropyridine of formula A-1, V is N(CH ₃ ) ₂ , OH, piperidine, morpholine, OMe, OEt or OiPr. Formatting A-2 then produces aldehyde A-3. Subsequent condensation with Ellman's sulfinamide gives the compound of formula A-4. Reduction and base-mediated cyclization of the compound of formula A-4 gives the compound of formula A-5. Alternatively, a compound of formula I, wherein R ¹ is cyclopropylmethyl, a compound of formula A-5. can be prepared using the synthetic method described for the preparation of intermediate 15c described herein. In the next step, an iridium-nickel mediated decarboxylative redox coupling under the influence of light between a chloropyridine of formula A-5 and a carboxylic acid of formula A-6 gives the compound of formula A-7. At this stage, the R ^3a substituent in A-6 must be the same group as is required in the final product of Formula I or a protected variant thereof. In addition, the substituent of R ⁷ and R ⁸ may be H, Boc, an alternative protecting group or alkyl; or, alternatively, the substituents R ^3a and R ⁸ may be part of a cyclic system, as desired in the final product of formula I. Coupling of a compound of formula A-7 with bromopyridine triazole A-8 in the presence of palladium or copper catalysis followed by removal of any protecting group(s) ) under standard conditions gives the pyrrolo[3,4-c]pyridin-1-one of formula I. At this stage, the R ^4N substituent in A-8 must be the same group as that required in the final product of formula I or a protected variant thereof .

Hydrazinolysis of bromopyridine ester A-9 (J. Med. Chem., 60(2), 722-748; 2017) followed by reaction of the corresponding hydrazide A-10 with dimethylformamide dimethyl acetal gives formamidine A-11. Condensation of formamidine A-11 with an amine gives triazole A-8.

Method B

Method B is the preparation of compounds of formula I wherein R ^3a and R ^3b are hydrogen. In the first step of Method B, from a compound of formula A-3 (V=N(CH ₃ ) ₂ , OH, piperidine, morpholine, OMe, OEt or OiPr), Negishi cross-coupling with the desired aminoalkyl zincate gives the compound of formula B-1. At this stage, the substituents R ⁷ and R ⁸ are either H or alkyl as represented in Formula I, or Boc, or an alternative protecting group. Subsequent condensation with Ellman's sulfonamide, reduction and base-mediated cyclization gives the compound of formula B-2. Coupling with bromopyridine triazole B-3 in the presence of palladium or copper catalysis and subsequent elimination of any protecting group(s) under standard conditions gives pyrrolo[3,4-c]pyridin-1-one of formula I. At this stage , the R ^4D substituent of Formula B-3 must be the same group as is required in the final product of Formula I or a protected variant thereof.

Pactam of formula B-4 is converted to the O-alkylimidate of formula B-5. At this stage, the R ^4D substituent of Formula B-4 must be the same group as is required in the final product of Formula I or a protected variant thereof. Cyclization of the O-alkylimidate of formula B-5 with the acylhydrazine of formula A-10 gives the compound of formula B-3.

Method C

Method C is the preparation of compounds of formula I wherein R ^3b is methyl. In the first step of Method C, palladium-mediated cross-coupling of a compound of formula A-5 and a boronic acid ester of formula C-1, followed by iron-mediated hydroazidation of the olefin or alkene, reduction and optional protection of the resulting amine, yields a compound of formula C-2. At this stage, the R ^3a substituent of formula C-1 and the R ¹ substituent of formula A-5 must be as presented in the desired product of formula I or a protected variant thereof. Cross-coupling of the compound of formula C-2 with bromopyridine triazole A-8 under palladium or copper catalysis followed by elimination of any protecting group(s) under standard conditions gives pyrrolo[3,4-c]pyridin-1-one of formula I. At this stage, the R ^4N substituent in A-8 must be the same group that is required in the R ⁴ of the final product of Formula I or a protected variant thereof.

Method D

Method D is the preparation of compounds of formula I wherein R ^3b is methyl. In the first step of Method D, the combination of a compound of formula C-2 with bromopyridine D-1 in the presence of copper or palladium catalysis produces a compound of formula D-2. At this stage, the substituents R ¹ , R ⁷ , R ⁸ and R ^3a C-2 and the substituent R ^4C D-1 must be represented by the same group that is required in the final product D-3 or a protected variant thereof. At this stage, R ⁷ and/or R ⁸ may also optionally be represented by protecting groups. Reaction of the compound of formula D-2 with the necessary amine gives the compound of formula D-3. At this stage, the R ^4N substituent must be the same group as is required in the final product, D-3, or a protected variant thereof. Cleavage of any protecting group(s) in the compound of formula D-3 ultimately yields the pyrrolo[3,4-c]pyridin-1-one of formula IA.

To prepare the compound of formula D-1, the compound of formula A-10 is reacted with the appropriate acyl chloride to yield the compound of formula D-4. At this stage, the R ^4C acyl chloride substituent is as represented in the final product of Formula I or a protected variant thereof. Cyclization of the compound of formula D-4 gives the compound of formula D-1.

Method E

Method E is the preparation of compounds of formula I wherein R ^3b is hydrogen. In the first stage of method E, either

i. Palladium-mediated cross-coupling between a compound of formula A-5 and tributyl(1-ethoxyvinyl)tin, followed by hydrolysis of the vinyl ester, gives the ketone of formula E-1, where the substituent R ^3a must be the same group as required in the final product of formula I, or

ii. Palladium-mediated cross-coupling between a compound of formula A-5 and zinc cyanide followed by reaction with an alkyl Grignard reagent produces a ketone of formula E-1, where the substituent R ^3a must be the same group required in the final product of formula I.

At this stage, the R ¹ substituent in A-5 must be the same group as is required in the final product of Formula I or a protected variant thereof.

From a compound of formula E-1, or

i. Reduction of the ketone followed by activation as a mesylate or chloride, replacement with an azide and reduction and optional protection gives a compound of formula A-7 (R ⁷ and R ⁸ are both H or R ⁷ is H and R ⁸ is Boc or an alternative protecting group), or

ii. Reduction of the ketone followed by activation as a mesylate or chloride, substitution with an alkylamine and optional protection provides a compound of formula A-7 (R ⁷ and/or R ⁸ is H, alkyl, Boc or an alternative protecting group), or

iii. Condensation with sulfinamide followed by reduction gives the compound of formula A-7 (R ⁷ is H and R ⁸ is SOtBu).

Coupling of the compound of formula A-7 with bromopyridine triazole A-8 in the presence of palladium or copper catalysis, followed by elimination of any protecting group(s) under standard conditions gives pyrrolo[3,4-c]pyridin-1-one from Formula I. At this stage, the R ^4N substituent in A-8 must be the same group as is required in the final product of Formula I or a protected variant thereof, within R ⁴ .

Method F

Method F is the preparation of compounds of formula I wherein R ^3b is hydrogen. In Method F, coupling a compound of Formula A-7 with bromopyridine triazole B-3 in the presence of palladium or copper catalysis followed by elimination of any protecting group(s) under standard conditions gives pyrrolo[3,4-c]pyridinium- 1-one from Formula I. At this stage, the R ^4C substituent in B-3 and the R ¹ , R ⁷ , R ⁸ and R ^3a substituents of Formula A-7 must be represented by the same group that is required in the final product of Formula I or its protected version. At this stage, the R ⁷ and/or R ⁸ substituents of formula A-7 may also be represented by Boc or another protecting group.

Method G

Method G is the preparation of compounds of formula I wherein R ^3b is hydrogen. In the first step of Method G, the bromopyridine of formula D-1 is reacted with the desired amine to produce the compound of formula G-1. At this stage, the substituents R ^4C and R ^4N must be the same group as required in the final product of Formula I or a protected variant thereof. Coupling of a compound of formula G-1 with compound A-7 in the presence of palladium or copper catalysis followed by elimination of any protecting group(s) under standard conditions gives pyrrolo[3,4-c]pyridin-1-one of formula I. At this stage, the substituents R ¹ , R ⁷ , R ⁸ and R ^3a of Formula A-7 must be the same group as is required in the final product of Formula I or a protected variant thereof. At this stage, the R ⁷ and/or R ⁸ substituents of formula A-7 may also be represented by Boc or another protecting group.

Method H

Method H is the preparation of compounds of formula I wherein R ^3b is methyl. In the first step of Method H, the combination of a compound of formula C-2 with bromopyridine triazole B-3 under palladium or copper catalysis followed by elimination of any protecting group(s) under standard conditions gives pyrrolo[3,4-c]pyridine -1-one of formula I. At this stage, the substituents R ¹ , R ⁷ , R ⁸ and R ^3a in C-2 and the substituent R ^4D in B-3 must be represented by the same group that is required in the final product of formula I or its protected version. R ⁷ and/or R ⁸ of formula C-2 may also optionally be represented by a protecting group at this stage.

Method I

Method I is the preparation of compounds of formula I wherein R ^3b is methyl. In the first step of Method I, coupling of a compound of formula C-2 with bromopyridine triazole G-1 in the presence of palladium or copper catalysis followed by elimination of any protecting group(s) under standard conditions gives pyrrolo[3,4-c] pyridin-1-one of formula I, where R ^3b represents methyl. At this stage, the Rl, R ⁷ , R ⁸ and R ^3a C-2 substituents and the R ^4C and R ^4N G-1 substituents must be the same group as is required in the final product, Formula I or a protected variant thereof. R ⁷ and/or R ⁸ of formula C-2 may also optionally be represented by a protecting group at this stage.

Method 7

Method 7 is the preparation of compounds of formula I wherein R ^3b is hydrogen. In the first step of process 7, the alcohol of formula 7-1 is protected to produce a compound of formula 7-2 (eg, X=OTBDMS). The compound of formula 7-2 is converted to the O-alkylimidate of formula 7-3. Cyclization of an O-alkylimidate of formula 7-3 with an acylhydrazine of formula A-10 gives the compound of formula 7-4. The compound of formula 7-4 is coupled with a compound of formula A-7 under a palladium or copper atmosphere, followed by removal of any protecting group(s) under standard conditions to yield pyrrolo[3,4-c]pyridine-1 -one from Formula I. At this stage, the substituents R ¹ , R ⁷ , R ⁸ and R ^3a in A-7 must be the same group as required in the final product of Formula I or a protected variant thereof. At this stage, the R ⁷ and/or R ⁸ substituents of formula A-7 may also optionally be represented by a protecting group.

Alternatively, to prepare compounds of formula I wherein R ⁴ is R ⁴ -ii and R ^4D is an F-substituted alkyl (X=fluorine) from a compound of formula 7-4 (X is a protected hydroxyl group), elimination The protecting group suggests a compound of formula 7-4, where X represents hydroxy. The alcohol can then be converted to an alkyl fluoride to give a compound of formula 7-4 wherein X is fluorine.

Method K

Method K is the preparation of compounds of formula I wherein R ^3a and R ^3b are hydrogen. In Method K, a compound of formula 7-4 (X is fluorine) is coupled with a compound of formula B-2 under palladium or copper catalysis, followed by removal of the protecting group(s) under standard conditions to give pyrrolo[3,4 -c]pyridin-1-one of formula I. At this stage, the substituents R ¹ , R ⁷ and R ⁸ in B-2 and the substituent X in formula 7-4 must be represented by the same group that is required in the final product of the formula I or its protected version. R ⁷ and/or R ⁸ of formula B-2 may also optionally be represented by a protecting group at this stage.

Method L

In the first step of Method L, the compound of formula L-1 is subjected to a Wittig olefination to produce the compound of formula L-2. Reduction and subsequent saponification of the compound of formula L-2 gives the compound of formula L-3. In these steps, the R ^4D substituent in L-1 and the R ^4D and R ^4F substituents in L-2 and L-3, as well as subsequent intermediates, are present in the final product of Formula I (as in R ⁴ ) or a protected variant thereof. Alternatively, certain compounds of Formula L-3 are commercially available. In the next step, the compound of formula L-3 is reacted with a compound of formula A-10 to produce the compound of formula L-4. Cyclization of the compound of formula L-4 gives the oxadiazole of formula L-5. Subsequent cyclization, preceded by thermal or acidic deprotection of the compound of formula L-5, gives the compound of formula L-6. A compound of formula L-6 is coupled with a compound of formula A-7, B-2 or C-2 in the presence of palladium or copper, followed by removal of any protecting group(s) under standard conditions to yield pyrrolo[3, 4-c]pyridin-1-one of formula I. The substituents R ¹ , R ² , R ^3a and R ^3b of formula I depend on their definitions in the previous methods for A-7, B-2 or C-2 or protected variants thereof.

Method M

Method M is the preparation of compounds of formula I wherein R ^3a and R ^3b are hydrogen. In Method M, the combination of the compound of formula B-2 with bromopyridine triazole A-8 under palladium or copper catalysis gives the protected pyrrolo[3,4-c]pyridin-1-one of formula I. At this stage, the substituents R ¹ , R ⁷ and R ⁸ of Formula B-2 and the R ^4N substituent of Formula A-8 must be the same group as required in the final product of Formula I, or a protected variant thereof, or as a protecting group (eg, R ⁷ =Boc). Cleavage of any protecting group(s) under standard conditions gives pyrrolo[3,4-c]pyridin-1-one of formula I.

Preparation of synthetic intermediates

For intermediates and examples prepared herein, where the stereochemistry is known, the stereochemistry is as shown and the name indicates the specific stereochemistry, such as (R) or (S). The stereochemistry reported herein is known because the compound was synthesized from known chiral starting materials or racemic mixtures, and the stereochemistry of certain examples or intermediates was confirmed using x-ray crystallography. In the case of the latter, the stereochemistry of other examples or intermediates will then be inferred based on the known chirality of the above examples or intermediates and the synthetic route. If the stereochemistry is unknown but the enantiomers are separated, "or1" or "or2" refers to the chiral carbon atom. In the name, a carbon with a resolved but not confirmed stereochemical center is designated by the symbol "ξ". The bond depicted at this carbon is a representation of stereochemistry; this means that the carbon will have this bond configuration (solid wedge) or the opposite configuration (shaded wedge). See, for example, example 6a and example 6b. Racemic compounds are indicated by the annotation “@1” and the bond is depicted as a specific stereochemistry, meaning that the carbon will have this bond configuration (solid wedge) and the opposite configuration (shaded wedge). See, for example, intermediate 15.

Intermediate 1: 2-bromo-6-(4-ethyl-4H-1,2,4-triazol-3-yl)pyridine

Step 1: 6-bromopyridine-2-carbohydrazide (1a)

To a solution of methyl 6-bromo-2-pyridinecarboxylate (16.0 g, 74.0 mmol) in MeOH (120 mL) was added hydrazine monohydrate (5.23 g, 88.8 mmol, 85%) and the mixture was stirred for 16 hours at room temperature. The resulting solution is concentrated to approximately half the volume and then triturated by adding 40 ml of methyl tert-butyl ether and stirring for 10 minutes. The resulting white solid was filtered and dried in vacuo to give the title compound (1a) (15 g, 94%). ^1H NMR (400 MHz, CDCl ₃ ) δ 8.82 (bs, 1H), 8.13 (dd, J=0.9, 7.5 Hz, 1H), 7.76-7.71 (m, 1H), 7.64 (dd, J=0.9, 8.0 Hz, 1H), 4.08 (shs, 2H). m/z (IER) for (C ₆ H ₆ BrN ₃ O) 217.5 (M+H) ⁺ .

Step 2: N'-[(6-bromopyridin-2-yl)carbonyl]-N,N-dimethylhydrazonoformamide (1b)

A solution of 6-bromopyridine-2-carbohydrazide (1a) (15.0 g, 69.4 mmol) in dimethylformamide dimethyl acetal (80 ml) was stirred at 80°C for 16 hours. The resulting mixture was concentrated under reduced pressure to obtain a residue. To this residue was added methyl tert-butyl ether (60 ml) and stirred for 40 minutes. The resulting yellow solid was filtered and dried to give the title compound (1b) (16 g, 85%). ^1H NMR (400 MHz, DMSO- _d6 ) δ 10.74 (s, 1H), 8.06 (s, 1H), 8.01-7.97 (m, 1H), 7.91 (t, J=7.7 Hz, 1H), 7.80 (dd, J=1.0, 7.8 Hz, 1H), 2.84 (s, 6H). m/z (ESI) for (C ₉ H ₁₁ BrN4O) 272.7 (M+H) ⁺ .

Stage 3: Intermediate 1

The flask was charged with N'-[(6-bromopyridin-2-yl)carbonyl]-N,N-dimethylhydrazonoformamide (1b) (2.0 g, 7.4 mmol), ethylamine (0.5 ml, 333 mg, 7 .4 mmol), acetic acid (3 ml) and MeCN (15 ml, 0.5 M). The solution is heated for 16 hours at 95°C. Add EtOAc (10 ml) and H ₂ O (10 ml). Add solid K ₂ CO ₃ until the pH of the aqueous layer becomes ~pH 8. The layers are separated and the aqueous layer is extracted with EtOAc (3×30 ml). The combined organic layers were washed with brine (30 ml), dried over Na ₂ SO ₄ , filtered and concentrated to dryness. The residue is suspended with EtOAc (0.3 ml) and petroleum ether (3 ml) for 5 min. The solids were collected by filtration to give Intermediate 1 (1.5 g, 80%) as a pale yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.75 (s, 1H), 8.19 (dd, J=7.7, 0.9 Hz, 1H), 7.99-7.90 ( m, 1H), 7.79 (dd, J=8.0, 0.9 Hz, 1H), 4.47 (kv, J=7.2 Hz, 2H), 1.38 (t, J=7 ,2 Hz, 3H); m/z (CIAD+) for (C ₉ H ₉ BrN ₄ ), 252.7 (M+H) ⁺ .

Intermediate 2: (5S)-3-(6-bromopyridin-2-yl)-5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole

Step 1: tert-butyl {(2S)-5-[2-(6-bromopyridin-2-carbonyl)hydrazinyl]-5-oxopentan-2-yl}carbamate (2a)

A solution of (4S)-4-[(tert-butoxycarbonyl)amino]pentanoic acid (600 mg, 2.76 mmol) in THF (13.8 mL, 0.2 M) was cooled to 0°C. A solution of propylphosphonic anhydride (50% solution in EtOAc, 3.62 mL, 6.08 mmol) was added to the solution at 0° C., after which the bath was removed and the reaction mixture was stirred for 30 minutes at room temperature. N,N-Diisopropylethylamine (2.89 mL, 16.6 mmol) and 6-bromopicolinohydrazide (656 mg, 3.04 mmol) were then added and the reaction mixture was stirred at room temperature for 22 hours. LCMS analysis showed consumption of starting material. The reaction is quenched with water (15 ml) and transferred to a separatory funnel with EtOAc (20 ml). The layers are separated and the organic phase is washed successively with 20% citric acid (20 ml), saturated NaHCO ₃ solution (20 ml) and brine (20 ml). The organic extract was then dried over MgSO ₄ , filtered and concentrated to dryness to give the title compound (2a) (1.07 g, 93% yield) as an off-white solid which was used without further purification. ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.76 (brs, 1H), 9.36 (brs, 1H), 8.15 (dd, J=0.9, 7.5 Hz, 1H), 7 .77-7.71 (m, 1H), 7.69-7.64 (m, 1H), 4.45 (shd, J=1.0 Hz, 1H), 3.91 (shs, 1H), 2.47-2.35 (m, 2H), 2.00-1.89 (m, 1H), 1.75-1.66 (m, 1H), 1.48 (s, 9H), 1. 22 (d, J=6.6 Hz, 3H). LCMS m/z (CIAD) for (C ₁₁ H ₁₅ BrN ₄ O ₂ ), 315.0 (M+H-Boc) ⁺ .

Step 2: tert-butyl {(2S)-4-[5-(6-bromopyridin-2-yl)-1,3,4-oxadiazol-2-yl]butan-2-yl}carbamate (2b)

To a solution of tert-butyl {[2S)-5-[2-(6-bromopyridin-2-carbonyl)hydrazinyl]-5-oxopentan-2-yl}carbamate (2a) (290 mg, 0.698 mmol) in DCM (2 .8 ml, 0.25 M) add triethylamine (0.292 ml, 2.09 mmol) and p-toluenesulfonyl chloride (160 mg, 0.838 mmol). The reaction mixture was stirred at room temperature for 16 hours. LCMS analysis showed consumption of starting material. Add ethylenediamine (0.047 mL, 0.698 mmol) to remove excess p-toluenesulfonyl chloride; upon addition, a precipitate immediately forms. After stirring at room temperature for 30 minutes, the reaction mixture was washed with 20% citric acid (5 ml) and the layers were separated. The aqueous layer was extracted with DCM (5 ml), then the combined organic layers were washed with brine (10 ml), dried over MgSO ₄ , filtered and concentrated to dryness to give the title compound (2b) (274 mg, 98% yield) as light yellow solid, which is used without further purification. ^1H NMR (400 MHz, CDCl ₃ ) δ 8.21 (dd, J=0.9, 7.6 Hz, 1H), 7.76-7.69 (m, 1H), 7.68-7, 65 (m, 1H), 4.39 (shs, 1H)), 3.85 (shs, 1H), 3.08-3.01 (m, 2H), 2.17-2.03 (m, 1H ), 2.03-1.90 (m, 1H), 1.44 (s, 9H), 1.22 (d, J=6.6 Hz, 3H). LCMS m/z (CIAD) for (C ₁₁ H ₁₃ BrN ₄ O), 297.0 (M+H-Boc) ⁺ .

Stage 3: Intermediate 2

Tert-butyl {(2S)-4-[5-(6-bromopyridin-2-yl)-1,3,4-oxadiazol-2-yl]butan-2-yl}carbamate (2b) is charged into a microwave-safe vessel ) (150 mg, 0.378 mmol) and trifluoroethanol (1.89 ml, 0.2 M) and sealed before heating in a microwave oven to 180 °C for 30 min. LCMS analysis showed consumption of starting material. The reaction mixture was concentrated and the residue was purified by flash chromatography (SiO ₂ , 100% heptane to 1:10 MeOH/EtOAc) to give intermediate 2 (74.3 mg, 71% yield) as a tan sticky solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.38 (d, J=7.7 Hz, 1H), 7.74 (t, J=7.8 Hz, 1H), 7.58 (d, J =7.9 Hz, 1H), 5.22-5.08 (m, 1H), 3.21-3.16 (m, 1H), 3.16-3.02 (m, 2H), 2, 55-2.44 (m, 1H), 1.60 (d, J=6.6 Hz, 3H). LCMS m/z (CIAD) for (C ₁₁ H ₁₁ BrN ₄ ), 279.1 (M+H) ⁺ . Determined as the single enantiomer SLC (10-60% methanol (0.5% NH ₃ ) in carbon dioxide at 400-450 bar, gradient time=2 min, flow rate=4 ml/min, Chiralpack IC-U 50 mm * 3 * mm column 1.6 µm). The stereochemistry of intermediate 2 is determined based on the use of (4S)-4-[(tert-butoxycarbonyl)amino]pentanoic acid in step 1.

The absolute configuration of intermediate 2 is unambiguously determined using small molecule X-ray crystallography. X-ray diffraction studies of single crystals are carried out on a Bruker APEX II Ultra CCD diffractometer equipped with Mo K _α radiation (λ=0.71073). The crystal is used as received (grown by vapor diffusion from dichloromethane). A colorless crystal measuring 0.200 x 0.075 x 0.060 mm is mounted on a Cryoloop with Paratone oil. Data are collected in a stream of nitrogen gas at 100(2) K) using φ and ω scans. The crystal-to-detector distance is 45 mm with an exposure time of 1 s (depending on the 2θ range) with a scan width of 0.80°. Data acquisition is completed at 100.0% to 25.242° in. θ. Data are integrated using Bruker SAINT software and scaled using SADABS software. Solving using direct methods (SHELXT) allows us to obtain a complete phase model corresponding to the proposed structure. All non-hydrogen atoms were refined anisotropically by the full-matrix least squares method (SHELXL-2014). All hydrogen atoms bonded to carbon are arranged using a "rider" model. Their positions are constrained relative to their parent atom using the appropriate HFIX command in SHELXL-2014. The positions of the hydrogen atoms CH, NH and OH are refined using the appropriate HFIX commands. Crystallographic data are summarized in Table 1.

Intermediate 3: (5S)-3-(6-bromopyridin-2-yl)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole

Step 1: tert-butyl{(2S)-1-[methoxy(methyl)amino]-1-oxobutan-2-yl}carbamate (3a)

A solution of (2S)-2-[(tert-butoxycarbonyl)amino]butanoic acid (2.0 g, 9.84 mmol) in THF (49.2 mL, 0.2 M) was cooled to 0°C. A solution of propylphosphonic anhydride (50% solution in EtOAc, 12.9 mL, 21.6 mmol) was added to the solution at 0° C. before removing the bath, and the reaction mixture was allowed to warm to RT and stirred for 30 minutes. N,N-Diisopropylethylamine (10.3 mL, 59.0 mmol) and methoxy(methyl)amine hydrochloride (1.06 g, 10.8 mmol) were then added and the reaction mixture was stirred at room temperature for 18 hours. LCMS analysis showed consumption of starting material. The reaction is quenched with water (40 ml) and transferred to a separatory funnel with EtOAc (40 ml). The layers are separated and the organic phase is washed successively with 20% citric acid (40 ml), saturated NaHCO ₃ solution (40 ml) and brine (40 ml). The organic extract was then dried over MgSO ₄ , filtered and concentrated to dryness to give the title compound (3a) (1.29 g, 53% yield) as a yellow oil which was used without further purification. ^1H NMR (400 MHz, CDCl ₃ ) δ 5.28-5.12 (m, 1H), 4.74-4.52 (m, 1H), 3.79 (s, 3H), 3.23 ( s, 3H), 1.85-1.73 (m, 1H), 1.64-1.54 (m, 1H), 1.46 (s, 9H), 0.96 (t, J=7, 5 Hz, 3H). LCMS m/z (CIAD) for (C ₁₁ H ₂₂ N ₂ O ₄ ), 247.1 (M+H) ⁺ .

Step 2: tert-butyl (S)-(1-oxobutan-2-yl)carbamate (3b)

A solution of tert-butyl {(2S)-1-[methoxy(methyl)amino]-1-oxobutan-2-yl} carbamate (3a) (1.29 g, 5.25 mmol) in THF (26.2 ml, 0.2 M) is cooled to 0°C. Lithium aluminum hydride (2.3 M solution in 2-MeTHF, 2.51 mL, 5.77 mmol) was added dropwise to the solution at 0°C. The reaction mixture is stirred for 40 minutes at 0°C. LCMS analysis showed consumption of starting material. The reaction is quenched with EtOAc (10 ml) and 1 M HCl (10 ml) and transferred to a separatory funnel. The layers are separated and the aqueous phase is extracted with EtOAc (3×10 ml). The combined organic extracts are then dried over MgSO ₄ , filtered and concentrated to dryness. The residue was purified by flash chromatography (SiO ₂ , 100% heptane to 100% EtOAc) to give the title compound (3b) (0.575 g, 59% yield) as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.60 (s, 1H), 5.09 (brs, 1H), 4.22 (brs, 1H), 2.04-1.90 (m, 1H) , 1.73-1.64 (m, 2H), 1.48 (s, 9H), 0.99 (t, J=7.5 Hz, 3H).

Step 3: ethyl (2E,AS)-4-[(tert-butoxycarbonyl)amino]hex-2-enoate (3c)

To a solution of tert-butyl (S)-(1-oxobutan-2-yl)carbamate (3b) (575 mg, 3.07 mmol) in DCM (6.14 mL, 0.5 M) add (ethoxycarbonylmethylene)triphenylphosphorane ( 1.60 g, 4.61 mmol). The reaction mixture was stirred at room temperature for 17 hours. LCMS analysis showed consumption of starting material. The reaction mixture was concentrated to dryness, then isopropanol (8.5 ml, 0.36 M) and zinc chloride (1.26 g, 9.21 mmol) were added to precipitate triphenylphosphine oxide. After stirring at room temperature for 30 minutes, the reaction mixture is filtered and concentrated to dryness. The residue was purified by flash chromatography (SiO ₂ , 100% heptane to 100% EtOAc) to give the title compound (3c) (653 mg, 83% yield, E/Z isomer ratio >20:1) as a colorless oil. ^1H NMR (400 MHz, CDCl ₃ ) δ 6.86 (dd, J=5.4, 15.6 Hz, 1H), 5.94 (dd, J=1.7, 15.7 Hz, 1H) , 4.50 (shs, 1H), 4.31-4.16 (m, 3H), 1.65-1.63 (m, 1H), 1.60-1.51 (m, 1H), 1 .47 (s, 9H), 1.32-1.30 (m, 3H), 0.99-0.95 (m, 3H). LCMS m/z (CIAD) for (C ₈ H ₁₅ NO ₂ ), 158.2 (M+H-Boc) ⁺ .

Step 4: ethyl (4S)-4-[(tert-butoxycarbonyl)amino]hexanoate (3d)

To a solution of ethyl ester (2E,4S)-4-[(tert-butoxycarbonyl)amino]hex-2-enoate (3c) (653 mg, 2.54 mmol) in methanol (12.7 ml, 0.2 M) add palladium on carbon (10% w/w 270 mg, 0.254 mmol). The reaction vessel is evacuated and refilled with H ₂ under dynamic vacuum for 10 seconds. Then the reaction mixture is stirred at room temperature under a pressure of 1 atm H ₂ for 18 hours. The mixture is stirred at room temperature for 17 hours. LCMS analysis showed consumption of starting material. The reaction mixture is filtered through Celite® and concentrated to dryness. The residue was purified by flash chromatography (SiO ₂ , 100% heptane to 100% EtOAc) to give the title compound (3d) (589 mg, 90% yield) as a colorless oil. ^1H NMR (400 MHz, CDCl ₃ ) δ 4.37-4.23 (m, 1H), 4.20-4.11 (m, 2H), 3.61-3.46 (m, 1H), 2.42-2.33 (m, 2H), 1.93-1.82 (m, 1H), 1.71-1.62 (m, 1H), 1.57-1.51 (m, 1H ), 1.46 (s, 9H), 1.44-1.38 (m, 1H), 1.30-1.26 (m, 3H), 0.94 (t, J=7.4 Hz, 3H).

Step 5: (4S)-4-[(tert-butoxycarbonyl)amino]hexanoic acid (3e)

To a solution of ethyl (4S)-4-[(tert-butoxycarbonyl)amino]hexanoate (3d) (589 mg, 2.27 mmol) in THF (11.4 ml, 0.2 M) and MeOH (5.7 ml , 0.4 M) add a solution of LiOH (544 mg, 22.7 mmol) in water (2.84 ml, 0.8 M). The reaction mixture was stirred at RT for 24 hours. The reaction mixture was concentrated before adding water and 1M HCl until pH 5 was reached. The aqueous layer was extracted with EtOAc (3×20 ml). The combined organic layers were washed with brine, dried over MgSO ₄ , filtered and concentrated to dryness to give the title compound (3e) (260 mg, 50% yield) as a light yellow oil, which was used without further purification.

Step 6: tert-butyl {(3S)-6-[2-(6-bromopyridin-2-carbonyl)hydrazinyl]-6-oxohexan-3-yl}carbamate (3f)

A solution of (4S)-4-[(tert-butoxycarbonyl)amino]hexanoic acid (3e) (260 mg, 1.12 mmol) in THF (5.6 mL, 0.2 M) was cooled to 0°C. A solution of propylphosphonic anhydride (50% solution in EtOAc, 1.47 mL, 2.47 mmol) was added to the solution at 0° C. before removing the bath, and the reaction mixture was stirred for 30 minutes at room temperature. N,N-Diisopropylethylamine (1.17 mL, 6.74 mmol) and 6-bromopicolinohydrazide (267 mg, 1.24 mmol) were then added and the reaction mixture was stirred at room temperature for 18 hours. The reaction is quenched with water (15 ml) and transferred to a separatory funnel with EtOAc (20 ml). The layers are separated and the organic phase is washed successively with 20% citric acid (20 ml), saturated NaHCO ₃ solution (20 ml) and brine (20 ml). The organic extract is then dried over MgSO ₄ , filtered and concentrated to dryness. The residue was purified by flash chromatography (SiO ₂ , 100% heptane to 100% EtOAc) to give the title compound (3f) (241 mg, 50% yield) as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.76 (brs, 1H), 9.29 (brs, 1H), 8.15 (d, J=6.8 Hz, 1H), 7.76-7 .71 (m, 1H), 7.68-7.64 (m, 1H), 4.45-4.31 (m, 1H), 3.69 (shs, 1H), 2.45-2.37 (m, 2H), 2.04-1.94 (m, 1H), 1.69-1.62 (m, 1H), 1.49 (s, 9H), 1.45 (shd, J=7 .3 Hz, 2H), 0.99 (t, J=1.5 Hz, 3H). LCMS m/z (CIAD) for (C ₁₂ H ₁₇ BrN ₄ O ₂ ), 329.0 (M+H-Boc) ⁺ .

Step 7: tert-butyl {(3S)-1-[5-(6-bromopyridin-2-yl)-1,3,4-oxadiazol-2-yl]pentan-3-yl}carbamate (3g)

To a solution of tert-butyl {( ₃₃ )-6-[2-(6-bromopyridin-2-carbonyl)hydrazinyl]-6-oxohexan-3-yl}carbamate (3f) (241 mg, 0.561 mmol) in DCM ( 2.2 ml, 0.25 M) add triethylamine (0.235 ml, 1.68 mmol) and p-toluenesulfonyl chloride (128 mg, 0.673 mmol). The reaction mixture was stirred at room temperature for 16 hours. LCMS analysis showed consumption of starting material. Add ethylenediamine (0.038 mL, 0.561 mmol) to remove excess p-toluenesulfonyl chloride; upon addition, a precipitate immediately forms. After stirring at room temperature for 30 minutes, the reaction mixture was washed with 20% citric acid (5 ml) and the layers were separated. The aqueous layer was extracted with DCM (5 ml), then the combined organic layers were washed with brine (10 ml), dried over MgSO ₄ , filtered and concentrated to dryness. The residue was purified by flash chromatography (SiO ₂ , 100% heptane to 100% EtOAc) to give the title compound (3g) (178 mg, 77% yield) as a white solid. ^1H NMR (400 MHz, CDCl ₃ ) δ 8.22 (dd, J=0.7, 7.6 Hz, 1H), 7.77-7.72 (m, 1H), 7.68-7, 65 (m, 1H), 4.35 (shd, J=8.2 Hz, 1H), 3.65 (shs, 1H), 3.07 (dt, J=6.1, 10.0 Hz, 2H ), 2.21-2.11 (m, 1H), 1.93-1.83 (m, 1H), 1.65-1.61 (m, 1H), 1.53-1.48 (m , 1H), 1.46 (s, 9H), 0.98 (t, J=7.4 Hz, 3H). LCMS m/z (CIAD) for (C ₁₂ H ₁₅ BrN ₄ O), 311.0 (M+H-Boc) ⁺ .

Stage 8: Intermediate 3

Load tert-butyl {(3S)-1-[5-(6-bromopyridin-2-yl)-1,3,4-oxadiazol-2-yl]pentan-3-yl}carbamate (3g) into a microwave-safe vessel ) (178 mg, 0.431 mmol) and trifluoroethanol (2.16 ml, 0.2 M) and sealed before heating in a microwave oven to 180 °C for 60 min. LCMS analysis showed consumption of starting material. The reaction mixture was concentrated and the residue was purified by flash chromatography (SiO ₂ , 100% heptane to 1:10 MeOH/EtOAc) to give intermediate 3 (116 mg, 91% yield) as a colorless oil. ^1H NMR (400 MHz, CDCl ₃ ) δ 8.26 (dd, J=0.1, 1.1 Hz, 1H), 7.67 (t, J=1.9 Hz, 1H), 7.49 (dd, J=0.1, 7.8 Hz, 1H), 4.90-4.83 (m, 1H), 3.07-2.89 (m, 3H), 2.58-2.49 (m, 1H), 2.04-1.97 (m, 1H), 1.83-1.70 (m, 1H), 0.96 (t, J=7.5 Hz, 3H). LCMS m/z (CIAD) for (C ₁₂ H ₁₃ BrN ₄ ), 293.0 (M+H) ⁺ . Determined 97.4% ee by SLC (10-60% methanol (0.5% NH ₃ ) in carbon dioxide at 400-450 bar, gradient time = 2 min, flow rate = 4 ml/min, Kromasil (R, R )Whelk-O column 50 mm * 3 mm * 1.8 µm). Stereochemistry is determined based on the use of (2S)-2-[(tert-butoxycarbonyl)amino]butanoic acid in the first step. The absolute configuration of intermediate 3 has been unambiguously established using small molecule X-ray crystallography.

Table 2 shows the results of crystalline X-ray diffraction carried out on a Bruker APEX II Ultra CCD diffractometer equipped with Mo Kα radiation (λ=0.71073). Crystals are used as received (grown by vapor diffusion from a mixture of ethyl acetate:methanol (10:1). Colorless crystals measuring 0.180 x 0.065 x 0.055 mm are placed on a Cryoloop with Paratone oil. Data are collected under a stream of nitrogen at 100 (2) K) using φ and ω scanning. The crystal-to-detector distance is 45 mm with an exposure time of 2 s (depending on the 2θ range) with a scan width of 0.80°. Data acquisition is 100.0% complete up to 25.242° in θ. Data are integrated using Bruker SAINT software and scaled using SADABS software. The solution using direct methods (SHELXT) allows us to obtain a complete phasing model corresponding to the proposed structure. All non-hydrogen atoms were refined anisotropically by the full-matrix least squares method (SHELXL-2014). All hydrogen atoms bonded to carbon are placed using the "rider" model. Their positions are constrained relative to their parent atom using the appropriate HFIX command in SHELXL-2014. The positions of the hydrogen atoms CH, NH and OH are refined using the appropriate HFIX commands.

Intermediate 4: (S,S)-2-methyl-N-[(1R)-1-{6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro- 1H-pyrrolo[3,4-c]pyridin-4-yl}ethyl]propane-2-sulfinamide

Step 1: {2-chloro-6-[(2R)-2-methylpyrrolidin-1-yl]pyridin-4-yl}(piperidin-1-yl)methanone (4a)

A solution of (2,6-dichloropyridin-4-yl) (piperidin-1-yl)methanone (600 mg, 2.32 mmol) and {2R)-2-methylpyrrolidine (591 mg, 6.95 mmol) in DMF (1 .5 ml) is stirred at 100°C for 16 hours. LCMS analysis showed consumption of starting material. The reaction mixture was cooled to room temperature, H ₂ O (40 ml) was added and the reaction mixture was extracted with DCM (3×40 ml). The combined organic layers are dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography (24 g SiO ₂ , 0-20% EtOAc/heptane) to give the title compound (4a) (664 mg, 93% yield). ^1H NMR (400 MHz, CDCl ₃ ) δ 6.44 (d, J=1.0 Hz, 1H), 6.21 (d, J=1.0 Hz, 1H), 4.12 (k, J =1.1 Hz, 1H), 3.72-3.62 (m, 2H), 3.54 (ddd, J=10.5, 7.6, 2.9 Hz, 1H), 3.40- 3.28 (m, 2H), 2.10-2.04 (m, 2H), 1.75-1.62 (m, 4H), 1.26 (t, J=7.2 Hz, 1H) , 1.21 (d, J=6.3 Hz, 2H); m/z (CIAD+) for (C ₁₆ H ₂₂ ClN ₃ O), 308.2 (M+H) ⁺ .

Step 2: 2-chloro-6-[{2R)-2-methylpyrrolidin-1-yl]-4-(piperidin-1-carbonyl)pyridine-3-carbaldehyde (4b)

To a solution of DMF (473 mg, 6.47 mmol) in DCM (3.0 ml) add POCl ₃ (992 mg, 6.47 mmol). The mixture is stirred for 10 min, and then a solution of {2-chloro-6-[(2R)-2-methylpyrrolidin-1-yl]pyridin-4-yl}(piperidin-1-yl)methanone (4a) (664) is added mg, 2.16 mmol) in DCM (3.0 ml). The mixture is stirred at reflux for 15 hours. LCMS analysis showed consumption of starting material. The reaction mixture is concentrated to dryness and slowly poured into saturated NaHCO ₃ solution (30 ml). The mixture is extracted with DCM (3×30 ml). The combined organic layers are dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography (24 g SiO ₂ , 0-40% EtOAc/heptane) to give the title compound (4b) (568 mg, 78% yield). ¹ H NMR (400 MHz, CDCl ₃ ) δ 10.07 (s, 1H), 5.99 (s, 1H), 4.13-4.65 (m, 1H), 3.68-3.83 ( m, 1H), 3.55-3.68 (m, 2H), 3.35-3.55 (m, 1H), 2.98-3.20 (m, 2H), 1.88-2, 17 (m, 3H), 1.71-1.83 (m, 2H), 1.55-1.67 (m, 3H), 1.46-1.55 (m, 1H), 1.31- 1.42 (m, 1H), 1.17-1.26 (m, 3H); m/z (CIAD+) for (C ₁₇ H ₂₂ ClN ₃ O ₂ ), 336.1 (M+H) ⁺ .

Step 3: N-[(E)-{2-chloro-6-[(2R)-2-methylpyrrolidin-1-yl]-4-(piperidin-1-carbonyl)pyridin-3-yl}methylidene]-2 -methylpropane-2-sulfinamide (4c)

Mixture of 2-chloro-6-[(2R)-2-methylpyrrolidin-1-yl]-4-(piperidin-1-carbonyl)pyridine-3-carbaldehyde (4b) (432 mg, 1.29 mmol), (R )-(+)-2-methyl-2-propanesulfinamide (187 mg, 1.54 mmol) and Ti(OEt) ₄ (880 mg, 3.86 mmol) in THF (10.0 ml) stirred at 45°C within 16 hours. LCMS analysis showed ~25% starting material remaining. Additional batches of (R)-(+)-2-methyl-2-propanesulfinamide (62.4 mg, 0.515 mmol) and Ti(OEt) ₄ (293 mg, 1.29 mmol) and the mixture was stirred at 50°C for 16 hours. LCMS analysis showed consumption of starting material. The reaction is cooled to RT. The mixture is diluted with DCM (50 ml) and washed with saturated NaHCO ₃ solution (35 ml) and brine (35 ml). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated to give the title compound (4c) (495 mg, 88% yield) as a white gum, which was used without further purification, m/z (CIAD+) for ( C ₂₁ H ₃₁ ClN ₄ O ₂ S), 440.2 (M+H) ⁺ .

Step 4: 4-chloro-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (4d)

Solution of N-[(E)-{2-chloro-6-[(2R)-2-methylpyrrolidin-1-yl]-4-(piperidin-1-carbonyl)pyridin-3-yl}methylidene]-2-methylpropane -2-sulfinamide (4 s) (495 mg, 1.13 mmol) in THF (15.0 ml) is cooled to 0°C and then a solution of LiBH ₄ (2.0 M in THF, 620 ml, 1. 24 mmol). The mixture is stirred at 0°C for 2 hours, then a solution of NaOMe (25% in MeOH, 2.5 ml, 10.1 mmol) is added. The reaction mixture is allowed to warm to room temperature and then stirred for 16 hours. The reaction mixture was diluted in DCM (60 ml) and washed with saturated aqueous NH ₄ Cl (60 ml) and brine (60 ml). The organic layer is dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography (SiO ₂ , 50-100% EtOAc/heptane) to give the title compound (4d) (199 mg, 70% yield) as a colorless foam. ^1H NMR (400 MHz, CDCl ₃ ) δ 6.68 (s, 1H), 6.45 (s, 1H), 4.35 (s, 2H), 4.21-4.14 (m, 1H) , 3.58 (ddd, J=10.5, 7.6, 2.8 Hz, 1H), 3.39 (kv, J=8.9 Hz, 1H), 2.13-1.97 (m , 2H), 1.75 (dt, J=5.2, 2.6 Hz, 1H), 1.23 (d, J=6.3 Hz, 3H). One hydrogen atom is assumed to be hidden by the water peak; m/z (CIAD+) for (C ₁₂ H ₁₄ ClN ₃ O), 252.3 (M+H) ⁺ .

Step 5: 6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-carbonitrile (4e)

Mixture of 4-chloro-6-[{2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (4d) (438 mg, 1 .74 mmol), Zn(CN) ₂ (306 mg, 2.6 mmol), DMF (15 ml) and Pd(PPh ₃ ) ₄ (100 mg, 0.09 mmol) are heated to 140 ° C in a microwave oven at for 30 minutes. The reaction mixture is diluted with DCM and filtered. The filter cake is washed with DCM. The filtrate was concentrated in vacuo and the crude compound was purified by flash chromatography (SiO ₂ , 0-100% EtOAc:DCM/heptane, 1:1) to give the title compound (4e) as a yellow solid (375 mg, 89% yield ). ^1H NMR (400 MHz, CDCl ₃ ) δ 7.00-6.94 (m, 1H), 6.46-6.32 (m, 1H), 4.55 (s, 2H), 4.29- 4.19 (m, 1H), 3.61 (ddd, J=2.6, 7.6, 10.3 Hz, 1H), 3.46-3.34 (m, 1H), 2.22- 2.02 (m, 3H), 1.86-1.72 (m, 1H), 1.25 (d, J=6.4 Hz, 3H); m/z (CIAD+) for (C ₁₃ H ₁₄ N ₄ O), 243.1 (M+H) ⁺ .

Step 6: 4-acetyl-6-[[2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (4f)

To a solution of 6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridine-4-carbonitrile (4e) (355 mg, 1.47 mmol) in THF (15 mL) and MeMgBr (4.88 mL, 14.7 mmol, 3.0 M in THF) was added in an ice/water bath. The resulting mixture is stirred at this temperature for 10 minutes, then warmed to room temperature and stirred for 2 hours. The mixture is quenched with 2 N HCl (6 ml) at 0°C and stirred at room temperature for 15 minutes. The mixture is neutralized with saturated NaHCO ₃ solution and extracted with DCM (3×40 ml). The combined organic layers are dried over Na ₂ SO ₄ , filtered and concentrated. The crude compound was purified by flash chromatography (SiO ₂ , 20-50% EtOAc/heptane) to give the title compound (4f) as a yellow solid (190 mg, 50% yield). ^1H NMR (400 MHz, CDCl ₃ ) 6.98 (s, 1H), 6.59 (brs, 1H), 4.70 (s, 2H), 4.28 (pcs, J=5.7 Hz, 1H), 3.64 (ddd, J=2.6, 7.5, 10.1 Hz, 1H), 3.48-3.38 (m, 1H), 2.70 (s, 3H), 2 ,19-2.10 (m, 2H), 2.08-2.01 (m, 1H), 1.79 (td, J=2.6, 5.0 Hz, 1H), 1.30 (d , J=6.2 Hz, 3H); m/z (CIAD+) for (C ₁₄ H ₁₇ N ₃ O ₂ ), 260.2 (M+H) ⁺ .

Stage 7: Intermediate 4

Add 4-acetyl-6-[{2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (4f) to a 40 ml bottle (200 mg, 0.77 mmol), (S)-tert-butylsulfinamide (187 mg, 1.54 mmol), THF (1.54 ml) and Ti(OEt) ₄ (1.54 mmol, 0.323 ml). The bottle is closed with a lid and heated to 80°C for 48 hours. The reaction mixture was then cooled to -78°C (dry ice/acetone bath) and L-selectride (1.54 mmol, 1.54 mL, 1.0 M in THF) was added dropwise. The resulting mixture was stirred at -78°C for 3 hours. The mixture is warmed to room temperature and quenched with saturated NH ₄ Cl (2 ml) dropwise, then add DCM (20 ml) and brine (20 ml). The mixture is filtered through Celite® and washed with DCM (40 ml). The organic layer was collected and the aqueous layer was extracted with DCM (20 ml). The combined organic layers are dried over Na ₂ SO ₄ , filtered and concentrated. The crude product is purified by flash chromatography (SiO ₂ solvent 0-10% MeOH in EtOAc) to give intermediate 4 as a yellow foam (127 mg, 45% yield) . m/z (CIAD+) for (C ₁₈ H ₂₈ N ₄ O ₂ S), 365.3 (M+H) ⁺ . Stereochemistry is determined based on the use of {2R)-2-methylpyrrolidine in step 1 and (S)-tert-butylsulfinamide in step 7.

Intermediate 5: (S,R)-2-methyl-N-[(1R)-1-{6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro- 1H-pyrrolo[3,4-c]pyridin-4-yl}propyl]propane-2-sulfinamide

Step 1: 6-[(2R)-2-methylpyrrolidin-1-yl]-4-propanoyl-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (5a)

Solution of 6-[{2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridine-4-carbonitrile (4e) (808 mg, 3 .33 mmol) in THF (8.0 ml) was cooled to 0°C and then treated with a solution of ethyl magnesium bromide (3.0 M in Et ₂ O, 11.1 ml, 33.3 mmol). The mixture is stirred at room temperature for 1 hour. LCMS analysis showed consumption of starting material. The reaction is cooled to 0°C and quenched by adding 2 N HCl. The mixture was stirred at room temperature for 10 minutes, neutralized by adding saturated NaHCO ₃ solution and then extracted with DCM (2×30 ml). The combined organic layers are dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography (24 g SiO ₂ , 0-100% EtOAc/heptane) to give the title compound (5a) (646 mg, 71% yield) as a yellow oil. ^1H NMR (400 MHz, CDCl ₃ ) δ 7.01 (s, 1H), 6.41 (brs, 1H), 4.73 (s, 2H), 4.39-4.22 (m, 1H) , 3.66 (ddd, J=2.6, 7.4, 10.1 Hz, 1H), 3.52-3.38 (m, 1H), 3.30-3.14 (m, 2H) , 2.26-2.10 (m, 2H), 2.06-2.01 (m, 1H), 1.81 (td, J=2.5, 5.1 Hz, 1H), 1.31 (d, J=6.2 Hz, 3H), 1.24 (t, J=7.3 Hz, 3H); m/z (CIAD+) for (C ₁₅ H ₁₉ N ₃ O ₂ ), 274.2 (M+H) ⁺ .

Step 2: (S,S)-2-methyl-N-[(1E)-1-{6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H -pyrrolo[3,4-c]pyridin-4-yl}propylidene]propane-2-sulfinamide (5b)

To a solution of 6-[(2R)-2-methylpyrrolidin-1-yl]-4-propanoyl-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (5a) (640 mg, 2.34 mmol) in THF (2.0 ml) add (S)-(-)-2-methyl-2-propanesulfinamide (568 mg, 4.68 mmol) and Ti (OEt) ₄ (2.14 g, 9.37 mmol). The mixture is stirred at 90°C for 23 hours. LCMS analysis showed consumption of starting material. The mixture is cooled to room temperature and then saline (40 ml) and DCM (30 ml) are added. The mixture is stirred for 10 minutes and then filtered through Celite®. The layers are separated. The aqueous layer was extracted with DCM (30 ml). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated to give the title compound (5b) (779 mg, 88% yield) as a white foam, which was directly used in the next step. m/z (CIAD+) for (C ₁₉ H ₂₈ N ₄ O ₂ S), 377.2 (M+H) ⁺ .

Stage 3: Intermediate 5

Solution of (S,S)-2-methyl-N-[(1E)-1-{6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H-pyrrolo [3,4-c]pyridin-4-yl}propylidene]propan-2-sulfinamide (5b) (654 mg, 1.40 mmol) in THF (12.0 ml) was cooled to -78°C and then treated dropwise a solution of L-selectride (1.0 M in THF, 2.5 ml, 2.5 mmol). The mixture is stirred at -78°C for 1.5 hours. Additional L-selectride (1.0 M, 0.417 mL, 0.417 mmol) was added and the mixture was stirred at -78°C for an additional 1.5 hours. The mixture was quenched with MeOH, diluted with brine (50 mL) and extracted with DCM (2 x 50 mL). The combined organic layers are dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography (12 g SiO ₂ , 0-10% MeOH/DCM) to give intermediate 5 (300 mg, 57% yield) as a yellow gum. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.66 (s, 1H), 6.48 (s, 1H), 5.14 (d, J=6.6 Hz, 1H), 4.35 (s, 2H), 4.27-4.14 (m, 2H), 3.59-3.44 (m, 1H), 3.29-3.18 (m, 1H), 2.14-2 .01 (m, 2H), 1.98-1.81 (m, 3H), 1.73-1.60 (m, 1H), 1.20 (d, J=6.2 Hz, 3H), 1.07 (s, 9H), 0.86-0.73 (m, 3H); m/z (CIAD+) for (C ₁₉ H ₃₀ N ₄ O ₂ S), 379.2 (M+H) ⁺ .

Stereochemistry is determined based on the use of (2R)-2-methylpyrrolidine in step 1 of the synthesis of intermediate 4 and (S)-(-)-2-methyl-2-propanesulfinamide in step 2. The absolute configuration of intermediate 5 is clearly established by X-ray diffraction analysis small molecules.

Table 3 presents X-ray diffraction data confirming the stereochemistry of intermediate 5. X-ray diffraction studies of single crystals are carried out on a Bruker Microstar APEX II CCD diffractometer equipped with Cu Kα radiation (λ=1.54178 Å). The absolute stereochemistry is determined finally (Flack=0.003 (12)). Crystals of intermediate 5 were grown from ether/pentane at 4°C. A piece of colorless crystal measuring 0.12 x 0.04 x 0.02 mm is mounted on a Cryoloop with Paratone oil. Data are collected in a stream of nitrogen gas at 100(2) K using ϕ and ω scans. The crystal-to-detector distance was 45 mm and exposure times were 4, 6, 8, 10, 14, or 20 seconds depending on the range 29 per frame at a scan width of 1.25°. Data acquisition is 97.0% complete up to 67500° in θ. Data are integrated using Bruker SAINT Software and scaled using SADABS software. Solving using direct methods (SHELXT) allows us to obtain a complete phase model corresponding to the proposed structure. All non-hydrogen atoms are refined anisotropically using the full-matrix least squares method (SHELXL-2014). All hydrogen atoms bonded to carbon are arranged using a "rider" model. Their positions are constrained relative to their parent atom using the appropriate HFIX command in SHELXL-2014.

Intermediate 6: 4-(2-aminopropan-2-yl)-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin- 1-he

To a suspension of 6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-carbonitrile (4e) (213 mg, 0.88 mmol) in THF (9 ml) at room temperature add a complex of bis(lithium chloride) and lanthanum (III) chloride (1.76 mmol, 2.93 ml, 0.6 M). The resulting mixture was stirred at room temperature for 30 minutes and then cooled to -78°C (dry ice/acetone bath) and methyl lithium (3.52 mmol, 2.20 mL, 1.6 M) was added dropwise. The resulting mixture was stirred at -78°C for 1 hour, then warmed to room temperature. The mixture is quenched with saturated NH ₄ Cl solution (20 ml), then brine (20 ml) and DCM (20 ml) are added. The mixture is stirred at room temperature for 5 minutes and filtered through Celite®. The organic layer was collected and the aqueous layer was extracted with DCM (20 ml). The combined organic layers are dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography (SiO ₂ , solvent 0-10% MeOH in DCM) to give intermediate 6 as a pale brown solid (157 mg, 65% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.60 (s, 1H), 6.42 (s, 1H), 4.56 (s, 2H), 4.17 (pcs, J=5, 7 Hz, 1H), 3.50 (ddd, J=2.4, 7.2, 9.8 Hz, 1H), 3.30-3.22 (m, 1H), 2.07-2.02 (m, 1H), 2.00-1.92 (m, 2H), 1.68 (td, J=2.4, 4.9 Hz, 1H), 1.41 (s, 3H), 1, 40 (s, 3H), 1.20 (d, J=6.2 Hz, 3H); m/z (CIAD+) for (C ₁₅ H ₂₂ N ₄ O), 275.2 (M+H) ⁺ .

Intermediate 7: (5R)-3-(6-bromopyridin-2-yl)-5-(fluoromethyl)-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole

Step 1: (5R)-5-(fluoromethyl)pyrrolidin-2-one (7a)

A solution of (5R)-5-(bromomethyl)pyrrolidin-2-one (600 mg, 3.37 mmol) in CH ₃ CN (6.74 ml, 0.5 M) was covered with aluminum foil to exclude light. To this solution was added a suspension of AgF (1.07 g, 8.43 mmol) in CH ₃ CN (12.5 ml, 0.27 M) in one portion. The reaction mixture was stirred at room temperature in the dark for 36 hours. The reaction mixture is filtered through Celite® and concentrated to dryness. The crude residue was purified by flash chromatography (SiO ₂ , 100% EtOAc to 1:1 EtOAc/acetone) to give the title compound (7a) (173 mg, 44% yield) as a colorless oil. ^1H NMR (400 MHz, CDCl ₃ ) δ 7.17 (brs, 1H), 4.49-4.17 (m, 2H), 4.01-3.84 (m, 1H), 2.44- 2.28 (m, 2H), 2.27-2.16 (m, 1H), 1.88-1.75 (m, 1H). ^19F NMR (376 MHz, CDCl ₃ ) δ-224.58 (s, 1F).

Step 2: tert-butyl (2R)-2-(fluoromethyl)-5-oxopyrrolidine-1-carboxylate (7b)

To a solution of (5R)-5-(fluoromethyl)pyrrolidin-2-one (7a) (170 mg, 1.45 mmol) in CH ₃ CN (6.74 ml, 0.5 M) add Boc ₂ O (348 mg , 1.60 mmol) and 4-(dimethylamino)pyridine (17.7 mg, 0.145 mmol). The reaction mixture is stirred at room temperature for 2 hours. TLC analysis (2:1 EtOAc:acetone) showed consumption of starting material. The reaction mixture was concentrated and purified by flash chromatography (SiO ₂ , 100% heptane to 100% EtOAc) to give the title compound (7b) (277 mg, 88% yield) as a colorless oil. ^1H NMR (400 MHz, CDCl ₃ ) δ 4.78-4.42 (m, 2H), 4.40-4.25 (m, 1H), 2.76-2.62 (m, 1H), 2.50-2.39 (m, 1H), 2.29-2.14 (m, 1H), 2.13-2.03 (m, 1H), 1.56 (s, 9H). ^19F NMR (376 MHz, CDCl ₃ ) δ-233.52 (s, 1F).

Step 3: tert-butyl[(2R)-1-fluoro-5-hydrazinyl-5-oxopentan-2-yl]carbamate (7c)

To a solution of tert-butyl (2R)-2-(fluoromethyl)-5-oxopyrrolidine-1-carboxylate (7b) (1.78 g, 8.19 mmol) in THF (41.0 mL, 0.2 M) hydrazine monohydrate (1.3 ml, 41.0 mmol) via syringe. The reaction mixture is stirred at room temperature for 30 minutes. TLC analysis (1:1 heptane:EtOAc) showed consumption of starting material. The reaction mixture was concentrated and then EtOAc (50 ml) and saturated aqueous NH ₄ Cl (50 ml) were added. The layers are separated and the aqueous layer is extracted with EtOAc (3×50 ml). The combined organic layers were dried over MgSO ₄ , filtered and concentrated to dryness to give the title compound (7c) (1.6 g, 78% yield) as a white solid, which was used without further purification. ^1H NMR (400 MHz, CDCl ₃ ) δ 5.15-5.00 (m, 1H), 4.52-4.44 (m, 1H), 4.39-4.33 (m, 1H), 3.93-3.75 (m, 1H), 2.40 (shs, 2H), 2.00-1.81 (m, 2H), 1.46 (s, 9H). ^19F NMR (376 MHz, CDCl ₃ ) δ-232.02 (s, 1F).

Step 4: tert-butyl {(2R)-5-[2-(6-bromopyridin-2-carbonyl)hydrazinyl]-1-fluoro-5-oxopentan-2-yl}carbamate (7d)

A solution of 6-bromopyridine-2-carboxylic acid (1.4 g, 6.93 mmol) in THF (34.7 ml, 0.2 M) was cooled to 0°C. A solution of propylphosphonic anhydride (50% solution in EtOAc, 9.1 ml, 15.2 mmol) was added to the solution at 0° C., after which the bath was removed and the reaction mixture was stirred for 30 minutes at room temperature. Then add N,N-diisopropylethylamine (7.24 ml, 41.6 mmol) and tert-butyl[(2R)-1-fluoro-5-hydrazinyl-5-oxopentan-2-yl]carbamate (7 s) (1 .90 g, 7.62 mmol), and the reaction mixture was stirred at room temperature for 17 hours. LCMS analysis showed consumption of starting material. The reaction is quenched with water (30 ml) and transferred to a separatory funnel with EtOAc (50 ml). The layers are separated and the organic phase is washed successively with 20% citric acid (30 ml), saturated NaHCO ₃ solution (30 ml) and brine (30 ml). The organic extract was then dried over MgSO ₄ , filtered and concentrated to dryness to give the title compound (7d) (2.37 g, 78% yield) as a yellow solid which was used without further purification. ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.74 (brs, 1H), 9.15 (brs, 1H), 8.14 (d, J=7.5 Hz, 1H), 7.77-7 .71 (m, 1H), 7.70-7.65 (m, 1H), 4.95 (shd, J=9.2 Hz, 1H), 4.62-4.36 (m, 2H), 4.12-3.98 (m, 1H), 2.49-2.37 (m, 2H), 2.03-1.95 (m, 2H), 1.49 (s, 9H). ^19F NMR (376 MHz, CDCl ₃ ) δ-232.01 (s, 1F). LCMS m/z (CIAD) for (C _n H ₁₄ BrFN ₄ O ₂ ), 333.1 (M+H-Boc) ⁺ .

Step 5: tert-butyl {(2R)-4-[5-(6-bromopyridin-2-yl)-1,3,4-oxadiazol-2-yl]-1-fluorobutan-2-yl}carbamate (7e )

To a solution of tert-butyl {(2R)-5-[2-(6-bromopyridin-2-carbonyl)hydrazinyl]-1-fluoro-5-oxopentan-2-yl}carbamate (7d) (2.3 g, 5 .3 mmol) in DCM (21.2 mL, 0.25 M), add triethylamine (2.22 mL, 15.9 mmol) and l-toluenesulfonyl chloride (1.21 g, 6.37 mmol). The reaction mixture is stirred at room temperature for 1.5 hours. LCMS analysis showed consumption of starting material. Ethylenediamine (0.355 mL, 5.31 mmol) was added to remove excess p-toluenesulfonyl chloride; a precipitate immediately formed during addition. After stirring at room temperature for 30 minutes, the reaction mixture was washed with 20% citric acid (20 ml) and the layers were separated. The aqueous layer was extracted with DCM (40 mL), then the combined organic layers were washed with brine (25 mL), dried over MgSO ₄ , filtered and concentrated to dryness to give the title compound (7e) (2.07 g, 94% yield). .) as a yellow solid, which is used without further purification. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.21 (d, J=1.6 Hz, 1H), 4.90-4.69 (m, 1H), 4.55 (bd, J=2, 8 Hz, 1H), 4.43 (shs, 1H), 4.08-3.84 (m, 1H), 3.15-3.05 (m, 2H), 2.29-2.06 (m , 2H), 1.45 (s, 9H). ^19F NMR (376 MHz, CDCl ₃ ) δ-231.82 (s, 1F). LCMS m/z (CIAD) for (C ₁₁ H ₁₂ BrFN ₄ O), 315.0 (M+H-Boc) ⁺ .

Stage 6: Intermediate 7

Tert-butyl {(2R)-4-[5-(6-bromopyridin-2-yl)-1,3,4-oxadiazol-2-yl]-1-fluorobutan-2-yl}carbamate ( 7e) (1.00 g, 2.41 mmol) and trifluoroethanol (10.4 ml, 0.17 M) and sealed before heating in the microwave to 180°C for 30 min. LCMS analysis showed consumption of starting material. The reaction mixture was concentrated and the residue was purified by flash chromatography (SiO ₂ , 100% heptane to 1:10 MeOH/EtOAc) to give intermediate 7 (564 mg, 78% yield) as a yellow oil. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.40 (d, J=7.3 Hz, 1H), 7.79-7.73 (m, 1H), 7.58 (d, J=1, 9 Hz, 1H), 5.26-5.01 (m, 2H), 4.89-4.71 (m, 1H), 3.25-3.08 (m, 3H), 2.96-2 .85 (m, 1H). ^19F NMR (376 MHz, CDCl ₃ ) δ-235.95 (s, 1F). LCMS m/z (CIAD) for (C ₁₁ H ₁₀ BrFN ₄ ), 297.0 (M+H) ⁺ . Stereochemistry is determined based on the use of (5R)-5-(bromomethyl)pyrrolidin-2-one in step 1.

Intermediate 8: tert-butyl methyl({6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4- yl}methyl)carbamate

Step 1: tert-butyl ({3-formyl-6-[(2R)-2-methylpyrrolidin-1-yl]-4-(piperidin-1-carbonyl)pyridin-2-yl}methyl)methylcarbamate (8a)

A solution of 2-chloro-6-[(2R)-2-methylpyrrolidin-1-yl]-4-(piperidin-1-carbonyl)pyridine-3-carbaldehyde (4b) (600 mg, 1.79 mmol) and PdCl ₂ (dppf) (261 mg, 0.357 mmol) in 1,4-dioxane (25.0 ml) was bubbled with N ₂ for 5 min and then heated to 80°C. A solution of {[(tert-butoxycarbonyl)(methyl)amino]methyl}(chlorido)zinc* (0.158 M in THF, 39.6 ml) was added at 80° C. and the mixture was stirred for a further 35 min at the same temperature. LCMS analysis showed consumption of starting material. The mixture is cooled to 30°C and filtered through Celite®. The filter cake is washed with DCM (5 x 10 ml) and the filtrate is concentrated to dryness. The residue is combined with the crude product obtained from a parallel reaction, identically with 100 mg of 2-chloro-6-[{2R)-2-methylpyrrolidin-1-yl]-4-(piperidin-1-carbonyl)pyridin-3- carbaldehyde. The mixture was purified by flash chromatography (SiO ₂ , 1:1 EtOAc/petroleum ether) to give the title compound (8a) (900 mg, 97% yield) as a yellow gum. ^1H NMR (400 MHz, DMSO- _d6 ) δ 9.84 (s, 1H), 6.40-6.06 (m, 1H), 4.91-4.45 (m, 2H), 4, 52-3.92 (m, 1H), 3.69-3.47 (m, 3H), 3.21-3.00 (m, 2H), 2.98-2.84 (m, 3H), 2.22-1.82 (m, 3H), 1.83-1.66 (m, 1H), 1.59 (s, 4H), 1.41 (s, 7H), 1.19 (d, J=18.2 Hz, 8H). m/z (IER+) for (C ₂₄ H ₃₆ N ₄ O ₄ ), 445.4 (M+H) ⁺ .

*as retrieved from: Angew. Chem. Inf. Ed. 2014, 53, 2678.

Step 2: tert-butyl methyl{[3-{{E)-[(2-methylpropan-2-sulfinyl)imino]methyl}-6-[(2R)-2-methylpyrrolidin-1-yl]-4-( piperidin-1-carbonyl)pyridin-2-yl]methyl}carbamate (8b)

Mixture of tert-butyl ({3-formyl-6-[(2R)-2-methylpyrrolidin-1-yl]-4-(piperidin-1-carbonyl)pyridin-2-yl}methyl)methylcarbamate (8a) (1, 40 g, 3.15 mmol), Ti (OEt) ₄ (1.44 g, 6.30 mmol) and (S)-{-)-2-methyl-2-propanesulfinamide (573 mg, 4.72 mmol) in THF (50.0 ml) is stirred at 50°C for 18 hours. Additional batches of Ti(OEt) ₄ (359 mg, 1.57 mmol) and (S)-(-)-2-methyl-2-propanesulfinamide (115 mg, 0.945 mmol) were added and the mixture was stirred at 50°C for additional 20 o'clock. LCMS analysis showed consumption of starting material. The reaction is quenched with saturated Na ₂ CO ₃ solution (150 ml) and DCM (100 ml) is added. The mixture is filtered through Celite® and the layers are separated. The aqueous layer was extracted with DCM (100 ml). The combined organic layers were washed with brine (150 mL), dried over Na ₂ SO ₄ , filtered and concentrated to give the title compound (8b) (1.7 g, >99%) as a yellow solid, m/ z (IER+) for (C ₂₈ H ₄₅ N ₅ O ₄ S), 548.5 (M+H) ⁺ .

Stage 5: Intermediate 8

To a solution of tert-butyl methyl{[3-{[E)-[(2-methylpropane-2-sulfinyl)imino]methyl}-6-[{2R)-2-methylpyrrolidin-1-yl]-4-(piperidine -1-carbonyl)pyridin-2-yl]methyl}carbamate (8b) (1.72 g, 3.41 mmol) in THF (20.0 ml) at 0°C add LiBH ₄ (68.6 mg, 3 .15 mmol). The reaction mixture is stirred at 0°C for 1 hour. TLC analysis showed consumption of starting material. The mixture is warmed to room temperature and a solution of NaOMe (30% in MeOH, 6.24 g, 34.6 mmol) is added. The mixture is stirred for 16 hours. LCMS analysis showed the formation of the desired mass of product. The reaction is concentrated to dryness. The residue was dissolved in EtOAc (40 ml) and washed with H ₂ O (40 ml). The aqueous layer was extracted with EtOAc (30 ml). The combined organic layers were washed with brine (60 mL), dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography (SiO ₂ , EtOAc) to give intermediate 8 (750 mg, 66% yield) as a white solid. ^1H NMR (400 MHz, CDCl ₃ ) δ 7.04-6.79 (m, 1H), 6.68 (s, 1H), 4.56-4.43 (m, 2H), 4.43- 4.33 (m, 2H), 4.27-4.16 (m, 1H), 3.58 (ddd, J=2.5, 7.3, 10.0 Hz, 1H), 3.45- 3.30 (m, 1H), 3.01-2.91 (m, 3H), 2.17-1.96 (m, 3H), 1.80-1.72 (m, 1H), 1. 53-1.37 (m, 9H), 1.26-1.23 (m, 3H); m/z (IER+) for (C ₁₉ H ₂₈ N ₄ O ₃ ), 361.2 (M+H) ⁺ .

Stereochemistry is determined based on the use of (2R)-2-methylpyrrolidine in Step 1 of the synthesis of intermediate 4.

Intermediate 9: tert-butylmethyl ({6-[methyl(propan-2-yl)amino]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl}methyl )carbamate

Step 1: 2-chloro-N,N-dimethyl-6-[methyl (propan-2-yl)amino]pyridin-4-carboxamide (9a)

A mixture of 2, 6-dichloro-N,N-dimethylpyridine-4-carboxamide (30.0 g, 137 mmol) and N-methylpropan-2-amine (50.1 g, 685 mmol) in MeCN (120 ml) was partitioned into three closed reaction vessels, and each was stirred at 100°C for 60 hours. LCMS analysis showed consumption of starting material. The reaction mixtures are combined and concentrated to dryness. The residue was purified by flash chromatography (SiO ₂ , 1:1 EtOAc/petroleum ether) to give the title compound (9a) (30.5 g, 87% yield) as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 6.45 (d, J=0.9 Hz, 1H), 6.31 (d, J=1.0 Hz, 1H), 4.82 (p, J =6.8 Hz, 1H), 3.08 (s, 3H), 2.97 (s, 3H), 2.83 (s, 3H), 1.16 (d, J=6.1 Hz, 6H ); m/z (IER+) for (C ₁₂ H ₁₈ ClN ₃ O), 255.9 (M+H) ⁺ .

Step 2: 2-chloro-3-formyl-N,N-dimethyl-6-[methyl (propan-2-yl)amino]pyridine-4-carboxamide (9b)

To a solution of DMF (21.9 g, 299 mmol) in EDC (120 ml) add POCl ₃ (45.9 g, 299 mmol) dropwise at 5-15°C. The mixture was stirred at room temperature for 15 min and 2-chloro-N,N-dimethyl-6-[methyl(propan-2-yl)amino]pyridin-4-carboxamide (9a) (25.5 g, 99. 7 mmol). The reaction mixture is stirred at 65°C for 16 hours. LCMS analysis showed consumption of starting material. The reaction mixture is cooled to room temperature and added dropwise to a saturated aqueous Na ₂ CO ₃ solution (900 ml). The mixture is extracted with DCM (2×300 ml). The combined organic layers are washed with brine (5×500 ml), dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography (SiO ₂ , 1:1 EtOAc/petroleum ether) to give the title compound (9b) (23.7 g, 84% yield) as a brown solid. ^1H NMR (400 MHz, CDCl ₃ ) δ 10.19 (s, 1H), 6.25 (brs, 1H), 3.12 (s, 3H), 3.02-2.85 (m, 3H) , 2.77 (s, 3H), 1.22 (shd, J=6.5 Hz, 6H); m/z (IER+) for (C ₁₃ H ₁₈ ClN ₃ O ₂ ), 283.9 (M+H) ⁺ .

Step 3: 2-chloro-N,N-dimethyl-3-{(E)-[(2-methylpropane-2-sulfinyl)imino]methyl}-6-[methyl(propan-2-yl)amino]pyridin- 4-carboxamide (9c)

Mixture of 2-chloro-3-formyl-N,N-dimethyl-6-[methyl (propan-2-yl)amino]pyridin-4-carboxamide (9b) (23.7 g, 83.5 mmol), (R )-(+)-2-methyl-2-propanesulfinamide (12.1 g, 100 mmol) and Ti(OEt) ₄ (38.1 g, 167 mmol) in THF (250 ml) were stirred at 50°C for 20 o'clock. LCMS analysis showed consumption of starting material. The reaction is concentrated to dryness. The residue is stirred with saturated NaHCO ₃ solution (300 ml) for 30 minutes. The mixture is filtered. The filter cake was washed with H ₂ O (3 x 80 mL) and petroleum ether (3 x 50 mL) and dried in vacuo to give the title compound (9c) (32.3 g, 99% yield) as a yellow solid. . ^1H NMR (400 MHz, DMSO-d6) δ 9.07 (s, 1H), 6.74 (s, 1H), 5.36-5.10 (m, 1H), 3.30 (s, 3H ), 3.26 (s, 3H), 3.10 (s, 3H), 1.54 (d, J=6.7 Hz, 6H), 1.49 (s, 9H); m/z (IER+) for (C ₁₇ H ₂₇ ClN ₄ O ₂ S), 387.2 (M+H) ⁺ .

Step 4: 4-chloro-6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (9d)

A solution of 2-chloro-N,N-dimethyl-3-{[E)-[(2-methylpropane-2-sulfinyl)imino]methyl}-6-[methyl(propan-2-yl)amino]pyridin-4- carboxamide (9 s) (32.3 g, 83.5 mmol) in THF (200 ml) was cooled to 0°C and LiBH ₄ (1.82 g, 83.5 mmol) was added. The mixture is stirred at room temperature for 1 hour. LCMS analysis showed consumption of starting material. NaOMe (165 g, 919 mmol) was added and the mixture was stirred at room temperature for 16 hours. The reaction mixture is filtered and the filter cake is washed with EtOAc (3×200 ml). The combined filtrate is concentrated to dryness. The residue is dissolved in DCM (300 ml) and washed with H ₂ O (500 ml). The aqueous layer was extracted with DCM (2×300 ml). The combined organic layers were washed with brine (500 ml), dried over Na ₂ SO ₄ , filtered and concentrated. The solid is suspended in a mixture of DCM (50 ml) and petroleum ether (120 ml) for 30 minutes. Solids are collected by filtration. The filter cake was dried in vacuo to give the title compound (9d) (11.3 g, 56% yield) as an off-white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.19 (s, 1H), 6.79 (s, 1H), 4.82 (p, J=6.7 Hz, 1H), 4.35 (d , J=1.2 Hz, 2H), 2.88 (s, 3H), 1.18 (d, J=6.7 Hz, 6H); m/z (IER+) for (C ₁₁ H ₁₄ ClN ₃ O), 239.9 (M+H) ⁺ .

Step 5: Methyl 6-[methyl(propan-2-yl)amino]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridine-4-carboxylate (9e)

Mixture of 4-chloro-6-[methyl (propan-2-yl) amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (9d) (11.3 g, 47 .1 mmol), PdCl ₂ (dppf) (2.16 g, 2.95 mmol) and TEA (14.3 g, 141 mmol) in MeOH (200 ml) stirred at 80°C for 40 hours in a CO atmosphere at 50 psi inch. TLC analysis (1:1 EtOAc/petroleum ether) showed consumption of starting material. The reaction is concentrated to dryness. The residue is dissolved in H ₂ O (200 ml) and extracted with DCM (2×150 ml). The combined organic layers were washed with brine (200 ml), dried over Na ₂ SO ₄ , filtered and concentrated. The residue was suspended in DCM (50 ml) for 30 minutes. Solids are collected by filtration. The filter cake is washed with petroleum ether (3×5 ml) and dried in vacuum. The filtrate is purified by flash chromatography (SiO ₂ , 40-70% EtOAc/DCM). The product containing fractions were concentrated to dryness and combined with the above filter cake to give the title compound (9e) (12.3 g, 99% yield) as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.12 (s, 1H), 6.79 (s, 1H), 4.89 (p, J=6.6 Hz, 1H), 4.68 (d , J=1.1 Hz, 2H), 3.97 (s, 3H), 2.96 (s, 3H), 1.21 (d, J=6.7 Hz, 6H); m/z (IER+) for (C ₁₃ H ₁₇ N ₃ O ₃ ), 263.9 (M+H) ⁺ .

Step 6: 4-(hydroxymethyl)-6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (9f)

To a mixture of methyl 6-[methyl(propan-2-yl)amino]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-carboxylate (9e) (1.0 g , 3.80 mmol) in THF (60 ml) add dropwise a solution of LiAlH ₄ (2.5 M in THF, 1.67 ml, 4.18 mmol) at 0°C. The mixture is stirred at 0°C for 1 hour, and then at 20°C for 16 hours. TLC analysis (1:1 EtOAc/petroleum ether) showed consumption of starting material. The mixture is quenched by adding 20% aqueous NaOH solution (0.5 ml). Na ₂ SO ₄ (4 g) is added to the mixture. The mixture was stirred for 30 minutes and then filtered. The filtrate was concentrated to dryness to give the title compound (9f) (890 mg, 99% yield) as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.04 (s, 1H), 6.82 (s, 1H), 4.85 (p, J=6.6 Hz, 1H), 4.67 (d , J=4.7 Hz, 2H), 4.33 (s, 2H), 4.09 (t, J=4.6 Hz, 1H), 2.92 (s, 3H), 1.20 (d , J=6.7 Hz, 6H); m/z (IER+) for (C ₁₂ H ₁₇ N ₃ O ₂ ), 236.0 (M+H) ⁺ .

Step 7: {6-[methyl(propan-2-yl)amino]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl}methylmethanesulfonate (9g)

To a mixture of 4-(hydroxymethyl)-6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (9f) (890 mg, 3.78 mmol) and TEA (957 mg, 9.46 mmol) in THF (20.0 ml) were added dropwise to MsCl (953 mg, 8.23 mmol) at 0° C. under N ₂ atmosphere. The mixture is stirred at 0°C. LCMS analysis showed consumption of starting material. The reaction mixture is diluted with saturated aqueous Na ₂ CO ₃ solution (30 ml) and extracted with EtOAc (2×20 ml). The combined organic layers were washed with brine (30 mL), dried over Na ₂ SO ₄ , filtered and concentrated to give the title compound (9g) (1.2 g, 99% yield) as a yellow solid, which was used without further purification, m/z (ESI+) for (C ₁₃ H ₁₉ N ₃ O ₄ S), 314.0 (M+H) ⁺ .

Step 8: 4-[(methylamino)methyl]-6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (9h)

To a mixture of {6-[methyl (propan-2-yl) amino]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl}methylmethanesulfonate (9g) (1, 18 g, 3.78 mmol) in THF (20.0 ml) add a solution of methylamine (2.0 M in THF, 37.8 ml, 75.6 mmol). The mixture is stirred for 1 hour. LCMS analysis showed consumption of starting material. The mixture was concentrated to dryness to give the title compound (9h) (940 mg, 99% yield) as a brown solid which was used without further purification. m/z (IER+) for (C ₁₃ H ₂₀ N ₄ O), 249.0 (M+H) ⁺ .

Stage 9: Intermediate 9

To a solution of 4-[(methylamino)methyl]-6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (9h) ( 940 mg, 11.4 mmol) and TEA (1.15 g, 11.4 mmol) in DCM (20.0 ml) add Boc ₂ 0 (1.65 mg, 7.57 mmol). The mixture is stirred for 30 minutes. LCMS analysis showed consumption of starting material. The reaction is concentrated to dryness. The residue was purified by flash chromatography (SiO ₂ , EtOAc) to give intermediate 9 (600 mg, 46% yield) as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 6.81 (s, 1H), 6.57-6.42 (m, 1H), 4.91 (p, J=6.6 Hz, 1H), 4 .48 (s, 2H), 4.35 (d, J=12.4 Hz, 2H), 2.92 (s, 3H), 2.88 (s, 3H), 1.48 (s, 5H) , 1.41 (s, 4H), 1.17 (d, J=6.7 Hz, 6H); m/z (IER+) for (C ₁₈ H ₂₈ N ₄ O ₃ ), 349.2 (M+H) ⁺ .

Intermediate 10: (S,S)-2-methyl-N-[(1R)-1-{6-[methyl(propan-2-yl)amino]-1-oxo-2,3-dihydro-1H- pyrrolo[3,4-c]pyridin-4-yl}propyl]propan-2-sulfinamide

Step 1: 6-[methyl(propan-2-yl)amino]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-carbonitrile

Mixture of 4-chloro-6-[methyl (propan-2-yl) amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (9d) (706 mg, 2.95 mmol), Zn(CN) ₂ (519 mg, 4.42 mmol), DMF (10 ml) and Pd(PPh ₃ ) ₄ (170 mg, 0.147 mmol) are heated to 140 °C in a microwave oven for 30 min. The reaction mixture is diluted with DCM and filtered. The filter cake is washed with DCM. The filtrate was concentrated in vacuo to give the title compound (10a) as a yellow solid (544 mg, 80% yield), which was used without further purification. ¹ H NMR (400 MHz, DMSO-d ₆ ) 9.01 (s, 1H), 7.13 (s, 1H), 4.80 (td, J=6.7, 13.4 Hz, 1H), 4.47 (s, 2H), 2.89 (s, 3H), 1.15 (d, J=6.7 Hz, 6H); m/z (CIAD+) for (C ₁₂ H ₁₄ N ₄ 0), 231.2 (M+H) ⁺ .

Step 2: 6-[methyl(propan-2-yl)amino]-4-propanoyl-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (10b)

Suspension of 6-[methyl(propan-2-yl)amino]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-carbonitrile (10a) (511 mg, 2.22 mmol) in THF (20.0 ml) is cooled to 0°C and then treated with a solution of ethyl magnesium bromide (3.0 M in Et ₂ O, 7.40 ml, 22.2 mmol). The mixture is stirred at room temperature for 1 hour. LCMS analysis showed consumption of starting material. The reaction mixture is cooled to 0°C and quenched by adding saturated NH ₄ Cl (60 ml), and then extracted with DCM (2×80 ml). The combined organic layers are dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography (24 g SiO ₂ , 0-100% EtOAc/heptane) to give the title compound (10b) (125 mg, 22% yield) as a yellow foam, m/z (CIAD+) for ( C ₁₄ H ₁₉ N ₃ O ₂ ), 262.2 (M+H) ⁺ .

Stage 3: Intermediate 10

To a solution of 6-[methyl(propan-2-yl)amino]-4-propanoyl-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (10b) (124 mg, 0.475 mmol ) in THF (6.0 ml) add (S)-(-)-2-methyl-2-propanesulfinamide (92 mg, 0.759 mmol) and Ti(OEt) ₄ (433 mg, 1.90 mmol). The mixture is stirred at reflux for 42 hours. LCMS analysis showed consumption of starting material. The mixture is cooled to -78°C and then treated dropwise with a solution of L-selectride (1.0 M in THF, 1.9 ml, 1.90 mmol). The mixture is stirred at -78°C for 4 hours. The mixture was quenched with MeOH, diluted with brine (30 mL) and extracted with DCM (20 x 2 mL). The combined organic layers are dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography (12 g SiO ₂ , 0-10% MeOH/DCM) to give intermediate 10 (72 mg, 41% yield) as a yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.67 (s, 1H), 6.64 (s, 1H), 5.25 (d, J=6.7 Hz, 1H), 4.94 (kv, J=6.7 Hz, 1H), 4.34 (s, 2H), 4.21 (kv, J=6.7 Hz, 1H), 2.84 (s, 3H), 1, 98-1.80 (m, 2H), 1.13 (dd, J=3.6, 6.7 Hz, 6H), 1.06 (s, 9H), 0.83 (t, J=1, 3 Hz, 3H). m/z (CIAD+) for (C ₁₈ H ₃₀ N ₄ O ₂ S), 367.2 (M+H) ⁺ . Stereochemistry is determined based on the use of (S)-(-)-2-methyl-2-propanesulfinamide in step 2.

Intermediate 11: tert-butyl{[6-(dimethylamino)-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl]methyl}methylcarbamate.

Step 1: 2-chloro-6-(dimethylamino)-N,N-dimethylpyridine-4-carboxamide (11a)

To a 3.0 L round bottom flask charged with methyl 2,6-dichloropyridine-4-carboxylate (58.0 g, 281 mmol) under nitrogen, add N,N-dimethylamine (38.1 g, 845 mmol) at 0 -10°C. Add THF (200 ml). A solution of i-PrMgCl (2.0 M in THF, 352 ml, 704 mmol) was added over 3 hours, maintaining the reaction temperature at 0-10°C. The reaction mixture is stirred for another 10 minutes at 0°C and then at 25°C for 18 hours. LCMS analysis showed consumption of starting material. The reaction is cooled in an ice bath and quenched by adding cold saturated aqueous NH ₄ Cl solution (500 ml), maintaining the reaction temperature <20°C. EtOAc (500 ml) was added and the layers were separated. The aqueous layer was extracted with EtOAc (500 ml). The combined organic layers are washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The resulting oil was taken up in heptane (200 ml) and concentrated on a rotary evaporator to form a solid. The suspension is stirred for 0.5 h and the solids are collected by filtration. The filter cake is washed with hexane (3×50 ml). The filter cake is suspended in EtOAc/petroleum ether 1:20 (100 ml) and the solids are collected by filtration. The filter cake was washed with 1:20 EtOAc/petroleum ether (3 x 30 mL) and then dried in vacuo to give the title compound (11a) (51 g, 80% yield) as a light yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 6.49 (d, J=0.7 Hz, 1H), 6.35 (d, J=0.9 Hz, 1H), 3.13-3.09 (m, 9H), 2.98 (s, 3H); m/z (IER+) for (C ₁₀ H ₁₄ ClN ₃ O), 227.9 (M+H) ⁺ .

Step 2: 2-chloro-6-(dimethylamino)-3-formyl-N,N-dimethylpyridine-4-carboxamide (11b)

Two series of reactions are carried out in parallel. POCl ₃ (85.9 g, 560 mmol) is added to a round bottom flask containing DMF (250 ml) at 15-25°C with stirring. The mixture was stirred at 15-25°C for 15 minutes and then 2-chloro-6-(dimethylamino)-N,N-dimethylpyridine-4-carboxamide (11a) (25.5 g, 112 mmol) was added. The mixture is stirred at 50°C for 16 hours. LCMS analysis showed consumption of starting material. The two reactions are combined and then quenched by slowly pouring into a cold aqueous saturated Na ₂ CO ₃ solution, maintaining a pH of ~9. The mixture is extracted with EtOAc (4×1.0 L). The combined organic layers are washed with brine (5×600 ml), dried over Na ₂ SO ₄ , filtered and concentrated. The crude product was combined with two additional reactions carried out in a similar manner with 7.5 g and 5.0 g of 2-chloro-6-(dimethylamino)-N,N-dimethylpyridine-4-carboxamide. The material is placed in EtOAc (200 ml) and suspended for 20 minutes. The suspension is filtered. The filter cake is washed with EtOAc (2×50 ml). The filter cake is suspended in petroleum ether/EtOAc (1:1) (80 ml) for 20 min. The suspension is filtered and the filter cake is washed with petroleum ether/EtOAc (1:1) (60 ml). The filter cake is dried under vacuum. The combined filtrate is concentrated to dryness. The residue is suspended in petroleum ether/EtOAc (100 ml) 1:1 for 30 min. The suspension is filtered and the filter cake is washed with petroleum ether/EtOAc (1:1) (2×50 ml) and dried in vacuo. The combined dried solids were suspended in petroleum solution (200 ml) for 10 minutes and collected by filtration. The filter cake is washed with petroleum ether (100 ml) and then concentrated in vacuo. The combined filtrate was concentrated under vacuum to ~50 ml and then left to stand for 2 days. The resulting solids were collected by filtration and the filter cake was washed with petroleum ether/EtOAc (3:2) (2×50 ml). The solids were combined with the title compound (11b) (52 g, 73% yield for combined set of reactions) as a yellow solid. ^1H NMR (400 MHz, CDCl ₃ ) δ 10.21 (d, J=0.6 Hz, 1H), 6.28 (d, J=0.6 Hz, 1H), 3.19 (s, 6H ), 3.13 (s, 3H), 2.77 (s, 3H); m/z (IER+) for (C ₁₁ H ₁₄ ClN ₃ O ₂ ), 255.9 (M+H) ⁺ .

Step 3: tert-butyl {[6-(dimethylamino)-4-(dimethylcarbamoyl)-3-formylpyridin-2-yl]methyl}methylcarbamate (11c)

A mixture of tert-butyl dimethyl carbamate (3.41 g, 23.5 mmol) and N,N,N,N-tetramethylenediamine (3.27 g, 28.2 mmol) in 135 ml THF was cooled to -55 °C under N ₂ . Slowly add a solution of s-BuLi (1.4 M in cyclohexane, 20.1 mL, 28.2 mmol), maintaining the solution temperature below -52°C (internal). The mixture was stirred for a further 30 minutes at -55°C and then treated with a solution of ZnCl ₂ (1.9 M in 2-methyltetrahydrofuran, 14.8 ml, 28.2 mmol), maintaining the reaction temperature below -52°C. The solution is stirred for an additional 40 minutes at -55°C, and then warmed to room temperature to obtain a solution of {[{tert-butoxycarbonyl)(methyl)amino]methyl}(chlorido)zinc (C=0.195 M). A portion of the pre-prepared zincate solution (90.2 mL, 17.6 mmol) was transferred to an oven-dried 250 mL round bottom flask under _N2 and concentrated to dryness to obtain a white foam. The flask is filled back with N ₂ . 2-chloro-6-(dimethylamino)-3-formyl-N,N-dimethylpyridine-4-carboxamide (11b) (3.0 g, 10 mmol), PdCl ₂ (dppf) (0.858 g, 1 .17 mmol), 1,4-dioxane (50 ml) and H ₂ O (0.159 g, 8.8 mmol). The suspension is transferred into the flask containing the zincate through a cannula, and then the mixture is stirred at 80°C for 80 minutes. LCMS indicated the formation of a mass of product with some starting material remaining. An additional aliquot of {[{tert-butoxycarbonyl)(methyl)amino]methyl}(chlorido)zinc solution (2.0 ml) was added and the mixture was stirred at 80°C for 20 minutes. No additional conversion is observed. The reaction is cooled to 0°C and quenched by adding a saturated aqueous solution of NH ₄ Cl (10 ml) and H ₂ O (20 ml). The mixture is stirred at 0°C for 20 minutes and then filtered through a pad of Celite®. The filtrate is extracted with EtOAc (4x). The combined organic layers are dried over Na ₂ SO ₄ , filtered and concentrated. The residue is purified by flash chromatography (80 g SiO ₂ , 0-100% EtOAc/heptane). The resulting white foam was triturated with MTBE and concentrated in vacuo to give the title compound (11c) (3.8 g, 95% yield) as a light yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.84 (d, J=6.8 Hz, 1H), 6.45 (s, 1H), 4.71 (s, 2H), 3.15 (s, 6H), 2.99 (s, 3H), 2.90 (s, 3H), 2.75 (s, 3H), 1.33 (d, J=69.8 Hz, 9H), m /z (IER+) for (C ₁₈ H ₂₈ N ₄ O ₄ ), 365.3 (M+H)+.

Step 4: tert-butyl {[6-(dimethylamino)-4-(dimethylcarbamoyl)-3-{(E)-[(2-methylpropan-2-sulfinyl)imino]methyl}pyridin-2-yl]methyl}methylcarbamate (11d)

To a solution of tert-butyl {[6-(dimethylamino)-4-(dimethylcarbamoyl)-3-formylpyridin-2-yl]methyl}methylcarbamate (11 c) (3.0 g, 8.0 mmol) and add (R) -(+)-2-methyl-2-propanesulfinamide (1.2 g, 9.88 mmol) in THF (40 ml) Ti(OEt) ₄ (5.63 g, 24.7 mmol). The mixture is stirred at 50°C overnight. The reaction mixture is cooled to room temperature, diluted with DCM (50 ml) and quenched by adding saturated NaHCO ₃ solution (20 ml). The solution is stirred vigorously for 20 minutes and then filtered through a pad of Celite®. Celite® is washed with DCM (3 times). The combined filtrate is dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography (80 g SiO ₂ , 0-100% EtOAc/heptane) to give the title compound (lid) (3.89 g, 97% yield) as a colorless foam. ^1H NMR (400 MHz, DMSO- _d6 ) δ 8.39 (s, 1H), 6.46 (s, 1H), 4.77-4.55 (m, 2H), 3.12 (s, 6H), 2.95 (s, 3H), 2.93 (s, 3H), 2.74 (s, 3H), 1.41 (s, 4H), 1.20 (s, 5H), 1. 12 (s, 9H); m/z (IER+) for (C ₂₂ H ₃₇ N ₅ O ₄ S), 468.4 (M+H) ⁺ .

Stage 5: Intermediate 11

Tert-butyl {[6-(dimethylamino)-4-(dimethylcarbamoyl)-3-{(E)-[(2-methylpropan-2-sulfinyl)imino]methyl}pyridin-2-yl]methyl} is loaded into a round-bottom flask. Methyl carbamate (11d) (3.89, 8.32 mmol) was added under N ₂ with THF (42 mL). The mixture is cooled to 0°C and then treated with a solution of NBH ₄ (2.0 M in THF, 4.37 ml, 8.73 mmol). The mixture was stirred at 0°C for 1 hour and then NaOMe solution (25% in MeOH, 17.1 mL, 74.9 mmol) was added at the same temperature. The reaction mixture is allowed to warm slowly to room temperature and stirred for 16 hours. LCMS analysis showed consumption of starting material. The mixture is diluted with DCM and washed with saturated aqueous NH ₄ Cl and brine. The organic layer is dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography (80 g SiO ₂ , 0-100% EtOAc/heptanes) to give intermediate 11 (1.7 g, 64% yield) as a colorless foam. ^1H NMR (400 MHz, DMSO- _d6 ) δ 8.68 (s, 1H), 6.68 (s, 1H), 4.42 (s, 2H), 4.23 (s, 2H), 3 .06 (s, 6H), 2.86 (s, 3H), 1.36 (m, 9H); LCMS m/z (ESI+) for (C ₁₆ H ₂₄ N ₄ O ₃ ), 321.2 (M+H) ⁺ .

Intermediate 12: 2-bromo-6-(4-propyl-4H-1,2,4-triazol-3-yl)pyridine.

A mixture of N'-[(6-bromopyridin-2-yl)carbonyl]-N,N-dimethylhydrazonoformamide (1b) (29.0 g, 106.8 mmol) and propan-1-amine (31.6 g, 534 mmol ) in MeCN (440 ml) and acetic acid (110 ml) were stirred at 95°C for 16 hours. LCMS analysis showed consumption of starting material. The reaction is concentrated to dryness. The residue is placed in H ₂ O (50 ml) and made alkaline to pH ~9 with 1 N NaOH (~500 ml). The mixture is extracted with EtOAc (3×150 ml). The combined organic layers were washed with brine (150 ml), dried over Na ₂ SO ₄ , filtered and concentrated. The residue is suspended with EtOAc (50 ml) for 10 minutes and the solid is collected by filtration. The filter cake was washed with petroleum ether (2 x 50 mL) and dried in vacuo to give the title compound (intermediate 12) (21.0 g, 74% yield) as a yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.74 (s, 1H), 8.20 (dd, J=0.7, 7.8 Hz, 1H), 7.98-7.91 ( m, 1H), 7.78 (dd, J=0.7, 8.0 Hz, 1H), 4.45-4.36 (m, 2H), 1.77 (sext, J=7.4 Hz , 2H), 0.87 (t, J=1.4 Hz, 3H); m/z (IER+) for (C ₁₀ H ₁₁ BrN ₄ ), 266.7 (M+H) ⁺ .

Intermediate 13: (S,S)-2-methyl-N-[(1R)-1-{6-[methyl(propan-2-yl)amino]-1-oxo-2,3-dihydro-1H- pyrrolo[3,4-c]pyridin-4-yl}ethyl]propan-2-sulfinamide

Step 1: 4-acetyl-6-(isopropyl(methyl)amino)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one

Methylmagnesium bromide (9.11 g, 76.4 mmol, 25.9 mL, 3.0 M) was added to a solution of 6-(isopropyl(methyl)amino)-1-oxo-2,3-dihydro-1H-pyrrolo[ 3,4-c]pyridine-4-carbonitrile (intermediate 10a) (1.76 g, 7.64 mmol) in THF (70 ml, c=0.11 M) at 0°C. The mixture was stirred at this temperature for 10 minutes, then allowed to warm to room temperature and stirred for 3 hours. The mixture is then quenched dropwise with 2 M HCl (7 ml) at room temperature and stirred for 30 minutes. Add saturated NaHCO ₃ solution (70 ml) to neutralize the solution and extract with DCM (3 x 100 ml). The combined organic layers are dried over Na ₂ SO ₄ and concentrated in vacuo. The resulting residue was purified by flash chromatography (40 g SiO ₂ , 10-40% EtOAc/heptane) to give the title compound (13a) as a yellow solid (553 mg, 29% yield). ^1H NMR (400 MHz, DMSO-d6) δ 8.82 (s, 1H) 7.06 (s, 1H) 4.88 (bd, J=6.12 Hz, 1H) 4.49 (s, 2H ) 2.94 (s, 3H) 2.61 (s, 3H) 1.19 (d, J=6.60 Hz, 6H); m/z (CIAD+) for (C ₁₃ H ₁₇ N ₃ O ₂ ), 248.2 (M+H) ⁺ .

Stage 2: Intermediate 13

Mixture of 4-acetyl-6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (13a) (8.10 g, 32 .75 mmol), (S)-tert-butylsulfinamide (7.94 g, 65.5 mmol) in Ti(OEt) ₄ (29.9 g, 131 mmol) in THF (100 ml) under N ₂ heated at 90°C for 72 hours and the reaction was monitored using LCMS. The reaction mixture is then cooled to 0°C and L-selectride (1 M in THF, 131 mmol, 131 ml) is added dropwise at 0°C. The mixture is stirred at 0°C for 3 hours. The mixture is quenched with saturated NH ₄ Cl (200 ml) and brine (200 ml) at 0-5°C. The suspension is filtered through a pad of Celite® and the filter cake is washed with DCM (500 ml). The filtrate layers were separated and the aqueous layer was extracted with DCM (2×100 ml). The combined organic layers were washed with brine (250 ml), dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography (SiO ₂ , 0-10% MeOH/DCM) to give intermediate 13 as a yellow solid (10 g, 87%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 6.83 (brs, 1H), 6.75 (s, 1H), 4.89-4.76 (m, 1H), 4.48-4.39 ( m, 1H), 4.37-4.26 (m, 3H), 2.83 (s, 3H), 1.55 (d, J=6.6 Hz, 3H), 1.19-1.07 (m, 15N). m/z (CIAD+) for (C ₁₇ H ₂₈ N ₄ O ₂ S), 353.2 (M+H) ⁺ . Stereochemistry is determined based on the use of (S)-tert-butyl sulfinamide in step 1.

Intermediate 14: (S,R)-2-methyl-N-[(1S)-1-{6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro- 1H-pyrrolo[3,4-c]pyridin-4-yl}ethyl]propane-2-sulfinamide

Mixture of 4-acetyl-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (4f) (4000 mg, 15 .43 mmol), (R)-2-methylpropane-2-sulfinamide (2240 mg, 18.5 mmol) in Ti(OEt) ₄ (20 ml) was stirred at 100° C. for 20 hours under N ₂ atmosphere. LCMS analysis showed consumption of starting material. The yellow solution is cooled to 0°C and diluted with THF (50 ml). Add L-selectride (38.6 mmol, 38.6 ml, 1M in THF) dropwise at 0°C. The mixture is then stirred at 10°C for 20 hours. LCMS showed remaining starting material. Additional L-selectride (38.6 mmol, 38.6 ml 1M in THF) was added dropwise at 0°C and the mixture was stirred at 10°C for an additional 2 hours. TLC analysis showed consumption of starting material. The mixture is quenched with a saturated aqueous solution of NH ₄ Cl (200 ml) at 0-5°C. The suspension is filtered through a pad of Celite® and the precipitate is washed with EtOAc (2×50 ml). The filtrate layers were separated and the aqueous layer was extracted with EtOAc (2×20 ml). The combined organic layers were washed with brine (30 mL), dried over Na ₂ SO ₄ , filtered and concentrated to leave a residue. The residue is first purified by flash chromatography (SiO ₂ , EtOAc/MeOH=10:1), and then further purified by preparative HPLC (ACSSH-CA; method column: YMC Triart C18 250*50 mm*7 µm, water (0.05% ammonium hydroxide v/v)-ACN to give intermediate 14 (2.5 g, 44%) as an off-white solid ¹ H NMR (400 MHz, CDCl ₃ ) δ 6.70 (s, 1H), 6.47 (shs, 1H), 4.57-4.34 (m, 4H), 4.20 (piece, J=5.9 Hz, 1H), 3.64-3.53 (m, 1H) , 3.47-3.35 (m, 1H), 2.20-1.99 (m, 3H), 1.77 (bdd, J=2.5, 4.7 Hz, 1H), 1.64 (d, J=6.5 Hz, 3H), 1.28 (d, J=6.2 Hz, 3H), 1.22 (s, 9H) for (C ₁₈ H ₂₈₎ . N ₄ O ₂ S), 365.1 (M+H) ⁺ Stereochemistry is determined based on the use of (R)-tert-butylsulfinamide.

Intermediate 15: tert-butyl {2-methoxy-1-[6-(1-methylcyclopropyl)-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl]ethyl }carbamate

Step 1: Ethyl 3-cyano-2-hydroxy-6-(1-methylcyclopropyl)pyridine-4-carboxylate (15a)

A mixture of 2-cyanoacetamide (10.0 g, 119 mmol) and TEA (12.0 g, 119 mmol) in EtOH (50 ml) was heated to 65 °C (internal temperature) until the solids dissolved, and then ethyl 3- (1-methylcyclopropyl)-3-oxopropanoate (24.6 g, 124 mmol). The mixture is stirred at 65°C for 2 hours. TLC analysis (1:10 EtOAc/petroleum ether) showed consumption of starting material. The reaction is cooled to 10°C. The resulting precipitate is collected by filtration. The filter cake is washed with MTBE (3×10 ml) and dried in vacuum. The filtrate is concentrated to dryness. EtOH (10 ml) was added and then MTBE (30 ml) was added. The resulting solids are collected by filtration. The filter cake is washed with EtOH (5 ml) and MTBE (2×10 ml) and dried in vacuum. The solids were combined to give the title compound (15a) (25.0 g, 85% yield) as a yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.74 (brs, 1H), 6.63 (brs, 1H), 4.36 (kv, J=7.1 Hz, 2H), 1.45 -1.27 (m, 6H)), 1.16-1.06 (m, 2H), 0.92-0.75 (m, 2H).

Step 2: Ethyl 2-chloro-3-cyano-6-(1-methylcyclopropyl)pyridine-4-carboxylate (15b)

To a solution of ethyl 3-cyano-2-hydroxy-6-(1-methylcyclopropyl)pyridine-4-carboxylate (15a) (24.0 g, 97.5 mmol) in MeCN (487 ml) add POCl ₃ (74.7 g, 487 mmol) dropwise at 30°C. The mixture is stirred at 65°C for 60 hours. TLC analysis (EtOAc) showed consumption of starting material. The solution is concentrated to remove residual POCl ₃ . The residue is poured onto ice and made alkaline with a saturated solution of NaHCO ₃ to pH ~8. The mixture is extracted with EtOAc (2×100 ml). The combined organic layers are dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by flash chromatography (1:10 EtOAc/petroleum ether) to give the title compound (15b) (21.9 g, 85% yield) as a light yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.79 (s, 1H), 4.50 (kv, J=7.2 Hz, 2H), 1.55 (s, 3H), 1.50-1 .40 (m, 5H), 1.03 (kV, J=3.9 Hz, 2H); m/z (IER+) for (C ₁₃ H ₁₃ ClN ₂ O ₂ ), 264.9 (M+H) ⁺ .

Step 3: 4-chloro-6-(1-methylcyclopropyl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (15 s)

To a solution of ethyl 2-chloro-3-cyano-6-(1-methylcyclopropyl)pyridine-4-carboxylate (15b) (2.5 g, 9.44 mmol) in EtOH (500 ml) add Raney Ni (2.0 g, 34.1 mmol). The black mixture is stirred at 30°C in an atmosphere of H ₂ at 30 psi. inch within 48 hours. TLC analysis (1:10 EtOAc/petroleum ether) showed consumption of starting material. The mixture is filtered through a layer of Celite®. The filter cake is washed with MeOH (250 ml). The combined filtrate is concentrated to dryness. The residue is suspended in EtOAc (5 ml) for 20 minutes and the suspension is filtered. The filter cake was washed with EtOAc (2 mL) and dried in vacuo to give the title compound (15 s) (1.1 g, 52% yield) as a gray solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.66 (brs, 1H), 7.23-7.05 (m, 1H), 4.56-4.34 (m, 2H), 1.55 ( shs, 3H), 1.39-1.11 (m, 2H), 0.99-0.66 (m, 2H); m/z (IER+) for (C ₁₁ H ₁₁ ClN ₂ O), 222.8 (M+H) ⁺ .

Stage 4: Intermediate 15

A 40 ml bottle is charged with 4-chloro-6-(1-methylcyclopropyl)-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (15 s) (200 mg, 0.898 mmol), Cs ₂ CO ₃ (644 mg, 1.98 mmol), N-(tert-butoxycarbonyl)-O-methylserine (394 mg, 1.80 mmol), NiCl ₂ ⋅glyme (39.5 mg, 0.18 mmol) , pyridin-2-yl-N-cyanoamidine (26.3 mg, 0.180 mmol), bis[2-(2,4-difluorophenyl)-5-methylpyridine]-4,40-di-tert-butyl-2 hexafluorophosphate, 20-bipyridine iridium(III) (18.2 mg, 0.018 mmol) and anhydrous DMF (27 ml). The mixture is bubbled with N ₂ for 2 minutes and irradiated with light 365 nM at 15-25°C for 18 hours (fan speed 5200 rpm, stirring speed 1200 rpm, 100% LED). The mixture is concentrated and purified by flash chromatography (SiO ₂ , 10-20% [0% MeOH in EtOAc] in petroleum ether) to give a yellow solid. Prep HPLC purification (column: YMC Triart C18 150*25 µm*5 µm, water (0.05% ammonium hydroxide v/v)-CAN) gives intermediate 15 as a white solid (315 mg, 32% ). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.65 (s, 1H), 6.77 (s, 1H), 5.66 (d, J=8.0 Hz, 1H), 4.94 (d , J=6.3 Hz, 1H), 4.63-4.48 (m, 2H), 3.77 (dd, J=8.9, 5.0 Hz, 1H), 3.58-3, 44 (m, 1H), 3.26 (s, 3H), 1.62 (s, 3H), 1.45 (s, 9H), 1.30 (kv, J=3.6 Hz, 2H), 0.88 (kV, J=3.4 Hz, 2H); m/z (IER+) for (C ₁₉ H ₂₇ N ₃ 0 ₄ ), 362.3 (M+H) ⁺ .

Examples:

The examples prepared herein have stereochemistry assigned based on the use of intermediates with confirmed stereochemistry, such as Examples 1 and 2, or based on the use of intermediates derived from stereospecific starting materials, such as Examples 3 and 4.

Example 1: 4-[(1R)-1-aminopropyl]-2-{6-[(5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2, 4]triazol-3-yl]pyridin-2-yl}-6-[{2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1 -He

Step 1: {S,S)-2-methyl-N-[(1R)-1-(2-{6-[(5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2,1 -c][1,2,4]triazol-3-yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H -pyrrolo[3,4-c]pyridin-4-yl)propyl]propane-2-sulfinamide

[

Mixture of (S,S)-2-methyl-N-[(1R)-1-{6-[{2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H-pyrrolo [3,4-c]pyridin-4-yl}propyl]propan-2-sulfinamide (intermediate 5) (63.0 mg, 0.17 mmol), (5S)-3-(6-bromopyridine-2- yl)-5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole (intermediate 2) (147 mg, 0.528 mmol), K ₃ PO ₄ (336 mg, 1.59 mmol), Pd ₂ (dba) ₃ (30.4 mg, 0.0528 mmol) and XantPhos (61.1 mg, 0.106 mmol) in 1,4-dioxane (4.5 ml, c= 0.1 M) is heated to 100°C. (block temperature) in a 40 ml bottle (closed) for 18 hours. The mixture is filtered through Celite®, washed with DCM. The filtrate was concentrated in vacuo and the crude compound was purified by flash chromatography (SiO ₂ , 0-10% MeOH/DCM) to give the title compound as a pale yellow foam (194 mg, 64% yield). m/z (CIAD+) for (C ₂₉ H ₄₀ N ₈ O ₂ S), 577.3 (M+H) ⁺ .

Stage 2: Example 1

A solution of 4 N HCl in 1,4-dioxane (0.01 mmol, 0.252 ml, 4 M) was added to the suspension of (S,S)-2-methyl-N-[(1R)-1-(2-{6- [(5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl]pyridin-2-yl}-6-[(2R )-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl)propyl]propan-2-sulfinamide (194 mg, 0.336 mmol ) in MeOH (12 ml). The resulting mixture was stirred at RT for 2 hours. Volatile substances were removed under reduced pressure. The crude product was purified by chiral SLC (Phenomenex Lux Cellulose-1 column 4.6×100 mm, 3 μm, 30% MeOH + 10 mM NH ₃ in CO ₂ at 120 bar, 4 ml/min) to obtain example 1 (146 mg, yield 92%), >99% di) as a pale yellow solid. ¹ H NMR (600 MHz, DMSO-d ₆ ) δ 8.48 (d, J=8.3 Hz, 1H), 8.01 (t, J=8.0 Hz, 1H), 7.91 (d , J=7.7 Hz, 1H), 6.67 (s, 1H), 5.22-5.03 (m, 3H), 4.33-4.21 (m, 2H), 3.60- 3.50 (m, 1H), 3.41-3.32 (m, 1H), 3.06-2.98 (m, 1H), 2.97-2.89 (m, 1H), 2. 88-2.76 (m, 1H), 2.37-2.27 (m, 1H), 2.09-1.86 (m, 5H), 1.69-1.62 (m, 1H), 1.44 (d, J=6.4 Hz, 3H), 1.15 (d, J=6.2 Hz, 3H), 0.86 (t, d=7.4 Hz, 3H); m/z (CIAD+) for (C ₂₆ H ₃₂ N ₈ O), 473.2 (M+H) ⁺ ; [α]D ₂₂ =+37.0° (c=0.1 M, MeOH).

Table 4 presents single crystal X-ray diffraction data confirming the stereochemistry of Example 1. X-ray diffraction studies of single crystals are carried out on a Bruker Microstar APEX II CCD diffractometer equipped with Cu K _α radiation (λ=1.54178 Å). Crystals of Example 1 were grown from acetonitrile/pentane. A piece of colorless crystal measuring 0.25 x 0.2 x 0.15 mm is mounted on a Cryoloop with Paratone oil. Data are collected in a stream of nitrogen gas at 100(2) K using ϕ and ω scans. The crystal-to-detector distance is 40 mm and the exposure time is 2 or 10 seconds depending on the 2θ range per frame at a scan width of 1.00°. Data acquisition is 99.4% complete to 67.679° by 9. Data is integrated using Bruker SAINT Software and scaled using SADABS software. Solving using direct methods (SHELXT) allows us to obtain a complete phase model corresponding to the proposed structure. All non-hydrogen atoms were refined anisotropically by the full-matrix least squares method (SHELXL-2014). All hydrogen atoms bonded to carbon are arranged using a "rider" model. Their positions are constrained relative to their parent atom using the appropriate HFIX command in SHELXL-2014.

Example 2: 4-[(1R)-1-aminoethyl]-2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2, 4]triazol-3-yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1 -He

Step 1: (S,S)-N-[{1R)-1-(2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][ 1,2,4]triazol-3-yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H-pyrrolo[3 ,4-c]pyridin-4-yl)ethyl]-2-methylpropane-2-sulfinamide

To a mixture of (S,S)-2-methyl-N-[(1R)-1-{6-[{2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H- pyrrolo[3,4-c]pyridin-4-yl}ethyl]propan-2-sulfinamide (intermediate 4) (50 mg, 0.14 mmol), {5S)-3-(6-bromopyridin-2-yl) )-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole (intermediate 3) (42.2 mg, 0.144 mmol) and K ₃ PO ₄ ( 87.4 mg, 0.412 mmol) in 1,4-dioxane (3 ml) add Pd ₂ (dba) ₃ (12.6 mg, 0.014 mmol) and XantPhos (15.9 mg, 0.027 mmol) under N ₂ atmosphere. After addition, the mixture is bubbled with N ₂ for 2 minutes. The resulting mixture is hermetically sealed and stirred at 85°C for 18 hours. The reaction mixture was concentrated in vacuo and the residue was purified by flash chromatography (SiO ₂ , 10% EtOAc/MeOH) to give the title compound (60 mg, 76% yield) as a yellow solid. ^1H NMR (400 MHz, CDCl ₃ ) δ 8.70 (d, J=8.2 Hz, 1H), 8.18 (d, J=1.4 Hz, 1H), 7.93 (t, J =8.0 Hz, 1H), 6.76 (s, 1H), 5.11-5.02 (m, 2H), 4.99-4.93 (m, 1H), 4.61-4, 49 (m, 2H), 4.34-4.27 (m, 1H), 3.67-3.57 (m, 1H), 3.46-3.35 (m, 1H), 3.11- 3.01 (m, 3H), 2.65 (td, J=3.7, 7.9 Hz, 1H), 2.19-2.08 (m, 3H), 2.07 (s, 1H) , 1.86-1.79 (m, 1H), 1.79 (bd, J=2.3 Hz, 1H), 1.69 (d, J=6.4 Hz, 3H), 1.31- 1.28 (m, 3H), 1.22 (s, 9H), 1.02 (t, J=7.5 Hz, 3H); m/z (IER+) for (C ₃₀ H ₄₀ N ₈ O ₂ S), δ 77.5 (M+H) ⁺ ;[α]D ₂₂ =+85.3° (c=0.1 M, MeOH).

Stage 2: Example 2

To a solution of (S,S)-N-[(1R)-1-(2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1 ,2,4]triazol-3-yl]pyridin-2-yl}-6-[{2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H-pyrrolo[3, 4-c]pyridin-4-yl)ethyl]-2-methylpropan-2-sulfinamide (60 mg, 0.10 mmol) in EtOAc (5 ml) is added dropwise with 4 M HCl in EtOAc (3 ml) at 0° WITH. The mixture is stirred at 20°C for 1 hour. The reaction mixture is concentrated in vacuo. EtOAc (10 ml) and water (10 ml) were added to the residue. The aqueous layer was made alkaline with saturated NaHCO ₃ solution and extracted with DCM (10 ml x 3). The combined organic layers were washed with brine (20 mL), dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was lyophilized for 16 hours to give Example 2 (47 mg, 95%) as a pale yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.59 (d, J=8.3 Hz, 1H), 8.09-8.02 (m, 1H), 8.02-7.97 ( m, 1H), 6.58 (s, 1H), 5.33 (d, J=17.1 Hz, 1H), 5.10 (d, J=17.1 Hz, 1H), 4.99 ( pcs., J=6.1 Hz, 1H), 4.29-4.19 (m, 1H), 4.12 (kv, J=6.9 Hz, 1H), 3.55 (pcs., J=8 .0 Hz, 1H), 3.04-2.85 (m, 3H), 2.58-2.55 (m, 1H), 2.11-1.96 (m, 4H), 1.79- 1.66 (m, 2H), 1.36 (d, J=6.5 Hz, 3H), 1.24-1.20 (m, 3H), 0.93 (t, J=7.4 Hz , 3H); m/z (IER+) for (C ₂₆ H ₃₂ N ₈ O), 473.4 (M+H) ⁺ .

Example 3: 4-[(1S)-1-aminoethyl]-2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2, 4]triazol-3-yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1 -He

Step 1: -N-[(1S)-1-(2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4 ]triazol-3-yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c] pyridin-4-yl)ethyl]-2-methylpropan-2-sulfinamide

To a suspension of (S,R)-2-methyl-N-[(1S)-1-{6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H- pyrrolo[3,4-c]pyridin-4-yl}ethyl]propan-2-sulfinamide (intermediate 14) (2000 mg, 5.487 mmol), (5S)-3-(6-bromopyridin-2-yl)- 5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole (intermediate 3) (1610 mg, 5.49 mmol) and _K3PO4 ( ₃₄₉₀ mg , 16.5 mmol) in 1,4-dioxane (30 ml), Pd ₂ (dba) ₃ (502 mg, 0.549 mmol) and XantPhos (635 mg, 1.10 mmol) were added under nitrogen. After addition, the mixture is bubbled with argon for 2 minutes. The resulting mixture is hermetically sealed and stirred at 85°C for 18 hours. TLC analysis showed consumption of starting material. The mixture is diluted with H ₂ O (100 ml) with stirring and the resulting suspension is filtered. The aqueous filtrate is separated. The solid is washed with DCM (100 ml). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated to give a yellow residue (3 g), which was purified by flash chromatography (SiO ₂ , 0-10% MeOH in EtOAc) to give a yellow solid. The solid was dissolved in DCM (50 ml) and silica-SH (4 g) was added. The yellow mixture is refluxed for 20 minutes. The mixture is filtered and the filter cake is washed with DCM/MeOH (10:1). The filtrate is concentrated. Treatment with silica-SH was repeated three more times to obtain the title compound (2.3 g, 72.7%) as a yellow solid. ^1H NMR (400 MHz, CDCl ₃ ) δ 8.71 (dd, J=0.7, 8.4 Hz, 1H), 8.18 (dd, J=0.1, 7.6 Hz, 1H) , 7.97-7.87 (m, 1H), 6.76 (s, 1H), 5.19-5.11 (m, 1H), 5.03-4.94 (m, 2H), 4 .65-4.57 (m, 1H), 4.57-4.51 (m, 1H), 4.24 (pcs, J=6.1 Hz), 1H), 3.66-3.56 ( m, 1H), 3.48-3.39 (m, 1H), 3.13-2.98 (m, 3H), 2.71-2.59 (m, 1H), 2.21-2, 03 (m, 4H), 1.87-1.75 (m, 2H), 1.68 (d, J=6.5 Hz, 4H), 1.30 (d, J=6.2 Hz, 3H ), 1.24 (s, 9H), 1.02 (t, J=1.5 Hz, 3H). m/z (IER) for (C ₃₀ H ₄₀ N ₈ O ₂ S), 577.5 (M+H) ⁺ .

Stage 2: Example 3

To a solution of (S,R)-N-[(1S)-1-(2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1 ,2,4]triazol-3-yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H-pyrrolo[3, 4-c]pyridin-4-yl)ethyl]-2-methylpropan-2-sulfinamide (2300 mg, 3.988 mmol) in EtOAc (10 ml) add 4 M HCl in EtOAc (20 ml) at 0°C. After addition, the mixture is stirred at 15°C for 1 hour. The reaction is monitored using LCMS. The resulting yellow suspension was concentrated to give a residue, which was dissolved in H ₂ O (30 ml) and extracted with EtOAc (25 ml). The aqueous layer was made alkaline with saturated NaHCO ₃ solution to pH ~8 and extracted with DCM (3×35 ml). The combined organic layers were washed with brine (20 ml), dried over Na ₂ SO ₄ , filtered and concentrated to give a solid, to which DCM (50 ml) was added. The solution is filtered through a GF/F glass microfiber filter (~0.7 µm) four times. The filtrate is concentrated to obtain a yellow solid, which is purified using pre-HPLC (column: Agela DuraShell C18 150 * 40 mm * 5 μm, water (0.05% ammonium hydroxide v/v) - CAN, gradient time 12 min , flow rate 25 ml/min). This gave Example 3 (1.5 g, 79.6%) as a yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.55 (d, J=8.0 Hz, 1H), 8.08-7.90 (m, 2H), 6.51 (s, 1H) , 5.26 (d, J=16.8 Hz, 1H), 5.08-4.92 (m, 2H), 4.23 (pcs, J=5.8 Hz, 1H), 4.09 ( kv, J=6.5 Hz, 1H), 3.53 (pcs, J=7.5 Hz, 1H), 3.30-3.23 (m, 1H), 3.07-2.83 (m , 3H), 2.62-2.53 (m, 1H), 2.16-1.85 (m, 6H), 1.77-1.64 (m, 2H), 1.33 (d, J =6.5 Hz, 3H), 1.21 (d, J=6.0 Hz, 3H), 0.93 (t, J=1.3 Hz, 3H). m/z (IER) for (C ₂₆ H ₃₂ N ₈ O), 473.4 (M+H) ⁺ .

Example 4: 4-[(1R)-1-aminoethyl]-2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2, 4]triazol-3-yl]pyridin-2-yl}-6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one

Step 1: (S,S)-N-[(1R)-1-(2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][ 1,2,4]triazol-3-yl]pyridin-2-yl}-6-[methyl(propan-2-yl)amino]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4 -c]pyridin-4-yl)ethyl]-2-methylpropane-2-sulfinamide

Solution of (S,S)-2-methyl-N-[(1R)-1-{6-[methyl(propan-2-yl)amino]-1-OKCO-2,3-dihydro-1H-pyrrolo[3 ,4-c]pyridin-4-yl}ethyl]propan-2-sulfinamide (intermediate 13) (627 mg, 1.78 mmol), (5S)-3-(6-bromopyridin-2-yl)-5 -ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole (intermediate 3) (521 mg, 1.78 mmol), K ₃ PO ₄ (1.13 g, 5.34 mmol), Pd ₂ (dba) ₃ (102 mg, 0.178 mmol) and XantPhos (206 mg, 0.356 mmol) in 1,4-dioxane (17.8 ml, c = 0.1 M) heated at 100°C in a 100 ml flask with a condenser under N ₂ for 18 hours. The mixture is cooled to rt, filtered and washed with DCM (20 ml). The filtrate is concentrated under reduced pressure. The residue was purified by flash chromatography (24 g, SiO ₂ , 0-10% MeOH/DCM) to give the title compound as a pale yellow solid (808 mg, 80%). m/z (CIAD+) for (C ₂₉ H ₄₀ N ₈ O ₂ S), 565.3 (M+H) ⁺ .

Stage 2: Example 4

A 4 N solution of HCl in 1,4-dioxane (1.07 mL, 4.29 mmol) was added to the solution of {S,S)-N-[(1R)-1-(2-{6-[{5S)- 5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl]pyridin-2-yl}-6-[methyl(propan-2-yl )amino]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl)ethyl]-2-methylpropan-2-sulfinamide (808 mg, 1.43 mmol) in MeOH (14.3 ml, c=0.1 M). The mixture is stirred at room temperature for 2 hours. Volatiles are removed under reduced pressure. The crude product was purified by chiral SLC (Phenomenex Lux Cellulose-1 column 21×250 mm, 30% MeOH + 10 mM NH ₃ in CO ₂ , maintained at 120 bar, 100 ml/min), to obtain example 4 (345 mg, 52% yield, >99% di, >99% pure) as a pale yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.56 (dd, J=1.0, 8.1 Hz, 1H), 8.07-8.01 (m, 1H), 8.00- 7.93 (m, 1H), 6.78 (s, 1H), 5.34-5.28 (m, 1H), 5.17-5.10 (m, 1H), 5.02-4, 95 (m, 1H), 4.89 (td, J=6.7, 13.4 Hz, 1H), 4.18 (kv, J=6.7 Hz, 1H), 3.01-2.94 (m, 2H), 2.93 (s, 3H), 2.62-2.54 (m, 2H), 2.09-1.97 (m, 1H), 1.84-1.72 (m , 1H), 1.41 (d, J=6.6 Hz, 3H), 1.20 (d, J=6.6 Hz, 6H), 0.92 (t, J=7.5 Hz, 3H ); m/z (CIAD+) for (C ₂₅ H ₃₂ N ₈ O), 461.3 (M+H) ⁺ . [α]D ₂₂ =+120.5° (c=0.1 M, MeOH).

Example 5: 4-[(1R)-1-aminopropyl]-2-{3-[(5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2, 4]triazol-3-yl]phenyl}-6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one

Step 1: (S,S)-2-methyl-N-[(1R)-1-(2-{3-[(5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2,1 -c][1,2,4]triazol-3-yl]phenyl}-6-[methyl(propan-2-yl)amino]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4 -c]pyridin-4-yl)propyl]propan-2-sulfinamide

A mixture of {S,S)-2-methyl-N-(1-{6-[methyl(propan-2-yl)amino]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-C ]pyridin-4-yl}propyl)propan-2-sulfinamide (intermediate 10) (63 mg, 0.17 mmol), (5S)-3-(3-bromophenyl)-5-methyl-6,7-dihydro -5H-pyrrolo[2,1-c][1,2,4]triazole (intermediate 2) (48 mg, 0.172 mmol), K ₃ PO ₄ (109 mg, 0.516 mmol), Pd ₂ (dba) ₃ (9.88 mg, 0.0172 mmol) and XantPhos (19.9 mg, 0.0344 mmol) in 1,4-dioxane (4.5 ml, c = 0.1 M) are heated at 100 ° C at 40 ml bottle (closed) for 18 hours. Volatiles are removed under reduced pressure. The residue was purified by flash chromatography (12 g, SiO ₂ , 0-10% MeOH/DCM) to give the title compound as a pale yellow foam (74.0 mg, 16%). m/z (CHI7AD+) for (C ₂₉ H ₄₀ N ₈ O ₂ S), 565.3 (M+H) ⁺ .

Stage 2: Example 5

A 4 N solution of HCl in 1,4-dioxane (0.164 ml, 0.655 mmol) was added to a solution of (S,S)-2-methyl-A7-[(1R)-1-(2-{3-[(5S)- 5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl]phenyl}-6-[methyl(propan-2-yl)amino]- 1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl)propyl]propan-2-sulfinamide (74 mg, 0.13 mmol) in MeOH (5.0 ml, c=0.026 M). The mixture is stirred at room temperature for 2 hours. Volatiles are removed under reduced pressure. The crude product is purified by chiral SLC (Phenomenex Lux Cellulose-1 column 4.6 x 100 mm, 3 µm, 5-60% MeOH + 10 mM NH ₃ in CO ₂ ramp for 3.0 minutes at 120 bar, 4 mL/min) to give Example 5 (11.6 mg, 19% yield, >99% di, >95% purity) as a pale yellow solid. ¹ H NMR (600 MHz, DMSO-d ₆ ) δ 8.54 (d, J=8.3 Hz, 1H), 8.35 (brs, 2H), 8.07 (t, J=8.0 Hz , 1H), 8.00-7.90 (m, 1H), 6.91 (d, J=0.9 Hz, 1H), 5.27-5.07 (m, 3H), 5.01 ( ws, 1H), 4.37 (ws, 1H), 3.14-2.97 (m, 2H), 2.94 (d, J=1.5 Hz, 3H), 2.92-2.85 (m, 1H), 2.42-2.35 (m, 1H), 2.06-1.91 (m, 2H), 1.50 (d, J=6.1 Hz), 3H), 1 .19 (d, J=6.6 Hz, 3H), 1.15 (d, J=6.6 Hz, 3H), 0.92 (t, J=7.3 Hz, 3H); m/z (CIAD+) for (C ₂₅ H ₃₂ N ₈ O), 461.3 (M+H) ⁺ . [α]D ₂₂ =+75.9° (c=0.2 M, MeOH).

Example 6: 4-[(1ξ)-1-amino-2-methoxyethyl]-6-(1-methylcyclopropyl)-2-{6-[(5S)-5-methyl-6,7-dihydro-5H-pyrrolo [2,1-c][1,2,4]triazol-3-yl]pyridin-2-yl}-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one

The stereochemistry of compounds 6a and 6b is depicted arbitrarily with the understanding that the two isomers have been separated (see description in previous section on intermediates).

Step 1: tert-butyl{2-methoxy-1-[6-(1-methylcyclopropyl)-2-{6-[(5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2,1- c][1,2,4]triazol-3-yl]pyridin-2-yl}-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl]ethyl} carbamate

A solution of tert-butyl{2-methoxy-1-[6-(1-methylcyclopropyl)-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl]ethyl}carbamate ( intermediate 15) (136 mg, 0.376 mmol), (5S)-3-(6-bromopyridin-2-yl)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1 ,2,4]triazole (intermediate 2) (105 mg, 0.376 mmol), K ₃ PO ₄ (240 mg, 1.13 mmol) and 1,4-dioxane (12 ml) were bubbled with nitrogen for 2 min. To this mixture were added Pd ₂ (dba) ₃ (34.5 mg, 0.038 mmol) and XantPhos (43.5 mg, 0.075 mmol). The reaction mixture is heated to 85°C and stirred at this temperature for 18 hours. The reaction mixture is cooled and combined with an identical 43 mg scale reaction run (intermediate 15). The combined mixture was concentrated and purified by flash chromatography (SiO ₂ , 10:1 EtOAc/MeOH) followed by prep. TLC (SiO ₂ , 10:1 EtOAc/MeOH) gave the title compound as a yellow solid (170 mg, 61%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.67 (dd, J=12.2, 8.3 Hz, 1H), 8.20-8.10 (m, 1H), 8.01-7, 84 (m, 1H), 5.66 (d, J=8.0 Hz, 1H), 5.35-5.11 (m, 3H), 5.03 (d, J=11.7 Hz, 1H ), 3.60-3.46 (m, 2H), 3.27 (d, J=8.5 Hz, 3H), 3.16-2.94 (m, 4H), 2.50 (d, J=8.7 Hz, 2H), 2.39 (s, 2H), 1.71 (s, 3H), 1.43 (s, 9H), 1.33 (kV, J=2.3 Hz, 2H), 0.92 (t, J=2.6 Hz, 2H). m/z (IER) for (C ₃₀ H ₃₇ N ₇ O ₄ ), 460.2 (M+H) ⁺ .

Stage 2: Example 6

To a solution of tert-butyl-{2-methoxy-1-[6-(1-methylcyclopropyl)-2-{6-[(5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2,1- c][1,2,4]triazol-3-yl]pyridin-2-yl}-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl]ethyl} carbamate (170 mg, 0.304 mmol) in DCM (5 ml) add dropwise 4M HCl in EtOAc (3 ml) at 0°C. The reaction mixture is stirred at 20°C for 1 hour and then concentrated. The resulting residue was diluted with H ₂ O (15 ml) and made alkaline to pH ~8 with saturated NaHCO ₃ solution. The mixture was extracted with DCM (3×8 ml) and the combined organic layer was washed with brine (20 ml), dried over Na ₂ SO ₄ , filtered and concentrated to give a yellow solid. The resulting solid is purified using prep. SLC (column: Daicel Chiralpak 250 mm * 30 mm * 10 µm, 0.1% NH ₃ ⋅H ₂ O EtOH) to obtain:

The isomer eluting first, as in example 6a (30 mg, 21%). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.56 (d, J=l, 9 Hz, 1H), 8.08 (t, J=7.8 Hz, 1H), 7.99 (d , J=1.8 Hz, 1H), 7.57 (s, 1H), 5.48 (d, J=18.1 Hz, 1H), 5.30 (d, J=18.1 Hz, 1H ), 5.12 (pcs, d=6.5 Hz, 1H), 4.30 (t, d=6.3 Hz, 1H), 3.65-3.52 (m, 2H), 3.30 -3.26 (m, 3H), 3.13-2.86 (m, 3H), 2.40 (bdd, J=8.4, 11.9 Hz, 3H), 1.59-1.52 (m, 6H), 1.31-1.24 (m, 2H), 0.89 (d, J=2.9 Hz, 2H). m/z (IER) for (C ₂₅ H ₂₉ N ₇ O), 560.3 (M+H) ⁺ . [α]D ₂₂ =+66.1° (c=0.1 M, MeOH).

The isomer eluting second as in example 6b (33 mg, 23%). ¹ H NMR (400 MHz, DMSO-d ₆ ) α 8.58 (d, J=7.9 Hz, 1H), 8.07 (t, J=7.9 Hz, 1H), 7.99 (d , J=7.3 Hz, 1H), 7.55 (s, 1H), 5.44 (d, J=18.1 Hz, 1H), 5.31 (d, J=18.1 Hz, 1H ), 5.16 (pcs, J=6.6 Hz, 1H), 4.29 (t, J=6.4 Hz, 1H), 3.55 (d, J=6.4 Hz, 2H), 3.25 (s, 3H), 3.13-2.86 (m, 3H), 2.40 (shdd, J=8.2, 11.6 Hz, 1H), 2.12 (shs, 2H) , 1.59-1.48 (m, 6H), 1.30-1.23 (m, 2H), 0.88 (d, J=3.l Hz, 2H). m/z (ESI) for (C ₂ 5H ₂₉ N ₇ O), 560.2 (M+H) ⁺ . [α]D ₂₂ =+41.5°.

Additional compounds of the present invention are prepared by modifications of the methods exemplified herein and are presented in Tables 5, 6, and 7. Unless otherwise indicated, all compounds having chiral carbon atoms are prepared and/or isolated as one enantiomer as shown in the structure. Chirality is further indicated by the name of the compound, which designates the specific stereochemistry as (R) or (S) for each chiral carbon. When all chiral carbon atoms are designated as known stereochemistry, the stereochemistry is based on the use of known chiral starting materials and/or confirmation of separated enantiomers by X-ray crystallography. Some compounds are prepared from racemic intermediates and separated into individual enantiomers by suitable chiral preparative SLC. If the stereochemistry is unknown but the enantiomers are separated, "or1" or "or2" refers to the chiral carbon atom. In the name, a carbon with a resolved but unconfirmed stereochemical center is indicated by the symbol. The bond drawn on that carbon is a representation of the stereochemistry; this means that the carbon will have this bond configuration (solid wedge) or the opposite configuration (shaded wedge). See, for example, Examples 101 and 102. As received, optical rotation (αD ₂₂ or αD ₂₀ ) is presented after the IUPAC name, represented simply as α.

Example 100: 4-(2-aminopropan-2-yl)-2-[6-(4-ethyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl]-6-[( 2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one

Mixture of 4-(2-aminopropan-2-yl)-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one (intermediate 6) (63.0 mg, 0.23 mmol), 2-bromo-6-(4-ethyl-4H-1,2,4-triazol-3-yl)pyridine (intermediate 1) (58 .1 mg, 0.230 mmol), K ₂ CO ₃ (69.8 mg, 0.505 mmol), Cul (10.9 mg, 0.058 mmol) and N,N-dimethylethylenediamine (0.115 mmol, 0.0125 ml) in CH ₃ CN (3.0 ml) is heated at 120°C with microwave irradiation for 30 min. The volatile material is removed and the residue is purified by flash chromatography (SiO ₂ , 0-10% MeOH/DCM). The product is further purified by HPLC (Phenemonex Gemini NX C18 column, 150×21.2 mm, 5 μm. Mobile medium A: water + 10 mM ammonium acetate, Mobile medium B: acetonitrile) to obtain example 100 as a pale yellow solid substances (49.3 mg, 49% yield). ¹ H NMR (600 MHz, DMSO-d ₆ ) δ 8.74 (s, 1H), 8.62 (d, J=8.4 Hz, 1H), 8.11-8.04 (m, 1H) , 8.03-7.90 (m, 1H), 6.55 (s, 1H), 5.40 (s, 2H), 4.65 (kv, J=7.2 Hz, 2H), 4, 24-4.19 (m, 1H), 3.29-3.33 (m, 2H, assumed; partially blocked by water peak), 2.16-2.03 (m, 2H), 1.98-1, 89 (m, 1H), 1.71-1.69 (m, 1H), 1.50 (t, J=7.l Hz, 3H), 1.47 (d, J=5.9 Hz, 6H ), 1.22 (d, J=6.2 Hz, 3H); m/z (CIAD+) for (C ₂₄ H ₃₀ N ₈ O), 447.3 (M+H) ⁺ .

Example 200: 2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl]pyridin-2- yl}-4-[(methylamino)methyl]-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one

Step 1: tert-butyl[(2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl ]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl) methyl]methylcarbamate

A mixture of tert-butyl methyl({6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl}methyl )carbamate (intermediate 8) (752 mg, 2.09 mmol), (5S)-3-(6-bromopyridin-2-yl)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1 -c][1,2,4]triazole (intermediate 3) (612 mg, 2.09 mmol), K ₃ PO ₄ (1.33 g, 6.26 mmol), Pd ₂ (dba) ₃ (120 mg, 0.209 mmol), and XantPhos (241 mg, 0.417 mmol) in 1,4-dioxane (20.9 ml, c=0.1 M) are heated at 100°C in a 100 ml condenser flask under nitrogen atmosphere for 18 hours. The mixture is cooled to rt, filtered and washed with DCM (20 ml). The filtrate is concentrated under reduced pressure. The residue was purified by flash chromatography (24 g, SiO ₂ , 0-10% MeOH/DCM) to give the title compound as a pale yellow solid (1.13 g, 85%). m/z (CIAD+) for (C ₃₁ H ₄₀ N ₈ O ₃ ), 573.30 (M+H) ⁺ .

Stage 2: Example 200

A 4 N solution of HCl in 1,4-dioxane (4.42 mL, 17.7 mmol) was added to the solution of tert-butyl[(2-{6-[(5S)-5-ethyl-6,7-dihydro-5H -pyrrolo[2,1-c][1,2,4]triazol-3-yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2 ,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl)methyl]methylcarbamate (1.01 g, 1.7 mmol) in DCM (22.1 ml, c=0.08 M) . The mixture is stirred at room temperature for 2 hours. Volatiles are removed under reduced pressure. The crude product was purified by SLC (ZymorSPHER HADP column with methanol and 10 mm NH ₃ ) to obtain the compound of example 200 as a pale yellow solid (640 mg, 77%). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.97 (bs, 1H), 8.60 (dd, J=0.9, 8.3 Hz, 1H), 8.17-8.06 ( m, 1H), 8.04-7.95 (m, 1H), 6.76 (s, 1H), 5.28 (d, J=16.6 Hz, 1H), 5.09-4.91 (m, 2H), 4.45-4.19 (m, 3H), 3.73-3.61 (m, 1H), 3.49-3.37 (m, 1H), 3.09-2 .87 (m, 3H), 2.76 (s, 3H), 2.61-2.54 (m, 1H), 2.15-1.96 (m, 4H), 1.80-1.64 (m, 2H), 1.21 (d, J=6.2 Hz, 3H), 0.93 (t, J=7.4 Hz, 3H). m/z (CIAD+) for (C ₂₆ H ₃₂ N ₈ O), 473.3 (M+H) ⁺ . [α]D ₂₂ =+76.0° (c=0.1M, MeOH).

Example 201: 2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl]pyridin-2- yl}-4-[(methylamino)methyl]-6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one

Step 1: tert-butyl[(2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl ]pyridin-2-yl}-6-[methyl(propan-2-yl)amino]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl)methyl] methyl carbamate

Add tert-butylmethyl({6-[methyl(propan-3-yl)amino]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c] to a screw cap flask equipped with a magnetic stirrer. pyridin-4-yl}methyl)carbamate (intermediate 9) (294 mg, 0.84 mmol), (5 S)-3-(6-bromopyridin-2-yl)-5-ethyl-6,7-dihydro -5H-pyrrolo[2,1-c][1,2,4]triazole (intermediate 3) (260 mg, 0.88 mmol), K ₃ PO ₄ (537 mg, 2.53 mmol), Pd ₂ (dba) ₃ (78.0 mg, 0.084 mmol), XantPhos (97.6 mg, 0.17 mmol) and 1,4-dioxane (16.9 ml, 0.05 M). The mixture is bubbled with N ₂ for 5 minutes before capping the flask and heating to 85°C for 22 hours. LCMS analysis showed consumption of starting material. The reaction mixture was concentrated and the residue was purified by flash chromatography (SiO ₂ , 100% heptane to 1:10 MeOH/EtOAc) to give the title compound (344 mg, 73% yield) as a yellow solid. ^1H NMR (400 MHz, CDCl ₃ ) δ=8.72-8.65 (m, 1H), 8.13 (d, J=7.5 Hz, 1H), 7.91 (t, J=7 .9 Hz, 1H), 6.89 (s, 1H), 5.13-4.85 (m, 4H), 4.25-4.08 (m, 1H), 3.10-2.96 ( m, 6H), 2.96-2.93 (m, 3H), 2.66-2.58 (m, 1H), 2.07-1.94 (m, 1H), 1.87-1. 74 (m, 1H), 1.53 (s, 1H), 1.50-1.35 (m, 9H), 1.23 (dd, J=2.8, 6.6 Hz, 6H), 1 .00-0.87 (m, 3H); LCMS m/z (CIAD) for (C ₃₀ H ₄₀ N ₈ O ₃ ), 561.3 (M+H) ⁺ .

Stage 2: Example 201

Into a flask equipped with a magnetic stirrer and containing tert-butyl[(2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4] triazol-3-yl]pyridin-2-yl}-6-[methyl(propan-2-yl)amino]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4 -yl)methyl]methylcarbamate (344 mg, 0.62 mmol) and CH ₂ Cl ₂ (12.3 ml, 0.05 M) add HCl (4 M in 1,4-dioxane, 1.53 ml, 6.2 mmol). After stirring at room temperature for 20 h, LCMS analysis showed consumption of the starting material. Toluene (5 ml) was added to the reaction flask and the reaction mixture was concentrated. The crude residue was passed through a strong ion exchange (SCX) column to remove residual HCl and concentrated to give Example 201 (278 mg, 98% yield) as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ=8.59 (d, J=7.8 Hz, 1H), 8.11-8.03 (m, 1H), 8.00-7.96 (m , 1H), 6.77 (s, 1H), 5.76 (s, 1H), 5.25 (s, 1H), 5.08 (s, 3H), 3.84 (s, 2H), 3 .01-2.88 (m, 6H), 2.61-2.54 (m, 1H), 2.39 (s, 3H), 2.05-1.93 (m, 1H), 1.80 -1.67 (m, 1H), 1.16 (d, J=6.7 Hz, 6H), 0.92 (t, J=7.4 Hz, 3H); LCMS m/z (CIAD) for (C ₂₅ H ₃₂ N ₈ O), 461.3 (M+H) ⁺ .

Example 202: 2-{6-[(5R)-5-(fluoromethyl)-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl]pyridin- 2-yl}-4-[(methylamino)methyl]-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1- He

Step 1: tert-butyl[(2-{6-[(5R)-5-(fluoromethyl)-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole-3 -yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4- yl)methyl]methylcarbamate

To a mixture of tert-butyl methyl({6-[(2R)-2-methylpyrrolidin-1-yl]-1-okco-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl} Methyl)carbamate (intermediate 8) (90 mg, 0.25 mmol), (5S)-3-(6-bromopyridin-2-yl)-5-ethyl-6,7-dihydro-5H-pyrrolo[2, 1-c][1,2,4]triazole (intermediate 7) (74.2 mg, 0.250 mmol) and K ₃ PO ₄ (159 mg, 0.749 mmol) in 1,4-dioxane (8 ml) add Pd ₂ (dba) ₃ (22.9 mg, 0.025 mmol) and XantPhos (28.9 mg, 0.0499 mmol) under N ₂ atmosphere. After addition, the mixture is bubbled with N ₂ for 2 minutes. The resulting mixture is hermetically sealed and stirred at 85°C for 18 hours. Saturated salt solution (30 ml) was added to the reaction mixture. The mixture is extracted with EtOAc (20 ml x 2). The combined organic layers are dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give a brown solid (250 mg). The material was purified by flash chromatography (SiO ₂ , DCM/EtOAc=1:1) to give the title compound (120 mg, 83%) as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.69 (d, J=8.3 Hz, 1H), 8.16 (d, J=7.6 Hz, 1H), 7.96-7.86 (m, 1H), 6.72 (s, 1H), 5.40 (d, J=30.1 Hz, 1H), 5.01 (dd, J=48.3, 16.6 Hz, 4H) , 4.71 (d, J=14.5 Hz, 1H), 4.26-4.05 (m, 1H), 3.59 (t, J=8.7 Hz, 1H), 3.39 ( kv, J=9.0, 8.4 Hz, 1H), 3.22-3.05 (m, 3H), 3.02 (s, 3H), 2.92-2.82 (m, 1H) , 2.19-1.96 (m, 4H), 1.77-1.71 (m, 1H), 1.43 (s, 9H), 1.26 (d, J=6.3 Hz, 3H ). LCMS m/z (ESI) for (C ₃₀ H ₃₇ FN ₈ O ₃ ), 577.2 (M+H) ⁺ .

Stage 2: Example 202

To a suspension of tert-butyl[(2-{6-[(5R)-5-(fluoromethyl)-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3- yl]pyridin-2-yl}-6-[{2R)-2-methylpyrrolidin-1-yl]-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl )methyl]methylcarbamate (120 mg, 0.208 mmol) in DCM (8 ml) was added dropwise with HCl in EtOAc (4 ml, 4 M) at 0°C. The mixture was stirred at room temperature for 2 hours and monitored by LCMS. The reaction mixture is then cooled to 0°C and HCl in EtOAc (5.0 mL, 4 M) is added. The mixture is stirred at room temperature for 30 minutes. The mixture is concentrated in vacuo to give a yellow solid (100 mg). Water (10 ml) was added to the residue and washed with EtOAc (10 ml). The aqueous layer was lyophilized for 16 hours to obtain Example 202 (94 mg, 88%, HCl salt) as a yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.21-9.00 (m, 2H), 8.61 (d, J=8.3 Hz, 1H), 8.14-8.08 ( m, 1H), 8.06-8.02 (m, 1H)), 6.75 (s, 1H), 5.43-5.29 (m, 1H), 5.23 (d, J=16 .6 Hz, 1H), 5.09-4.91 (m, 3H), 4.40-4.29 (m, 3H), 3.70-3.59 (m, 1H), 3.13- 2.92 (m, 3H), 2.79-2.71 (m, 4H), 2.15-1.96 (m, 3H), 1.77-1.70 (m, 1H), 1. 21 (d, J=6.3 Hz, 3H). LCMS m/z (ESI) for (C ₂₅ H ₂₉ FN ₈ O), 477.4 (M+H) ⁺ .

Example 300: 6-(dimethylamino)-4-[(methylamine-)methyl]-2-[6-(4-propyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl]- 2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one

Step 1: tert-butyl({6-(dimethylamino)-1-oxo-2-[6-(4-propyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl]-2 ,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl}methyl)methylcarbamate

Tert-Butyl-{[6-(dimethylamino)-1-oxo-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-4-yl]methyl}methylcarbamate mixture (intermediate 11) (1, 33 g, 4.15 mmol), 2-bromo-6-(4-propyl-4H-1,2,4-triazol-3-yl)pyridine (intermediate 12) (1.16 g, 4.36 mmol ), Pd ₂ (dba) ₃ (380 mg, 0.415 mmol), XantPhos (480 g, 0.830 mmol) and K ₃ PO ₄ (2.64 g, 12.5 mmol) in 1,4-dioxane (46 ml) degassed with nitrogen for 5 minutes, and then stirred at 85°C for 16 hours. The reaction was analyzed by LCMS, which showed the consumption of starting material. The mixture is cooled to room temperature, filtered through a pad of Celite® and concentrated in vacuo. The residue is suspended with EtOAc (15 ml) for 10 min and the solids are collected by filtration. The filter cake is washed with EtOAc (4x) and then dried in vacuo. The residue is purified by flash chromatography (40 g SiO ₂ , 0-100% EtOAc/heptane, then 10% MeOH/EtOAc) to give a light yellow solid. The material is dissolved in a mixture of 1:9 EtOH/DCM and treated with ultrapure Si-ThioSiO ₂ (1.59 g). The mixture is stirred for 2 hours and filtered. The filter cake is washed with 1:9 EtOH/DCM and the combined filtrate is concentrated in vacuo. The residue was dissolved in 1:9 EtOH/DCM and treated with ultrapure Si-ThioSiO ₂ (1.32 g). The mixture is stirred for 3 hours and then filtered. The filter cake is washed with 1:9 EtOH/DCM and the combined filtrate is concentrated. The residue is dissolved in 1:9 EtOH and treated with ultrapure Si-ThioSiO ₂ (1.22 g). The mixture is stirred for 16 hours and then filtered. The filter cake is washed with a 1:9 EtOH/DCM mixture. The combined filtrate was concentrated to dryness to give the title compound (2.08 g, 95% yield) as a light yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.74 (s, 1H), 8.61 (d, J=8.4 Hz, 1H), 8.09 (t, J=8.0 Hz , 1H), 7.97 (d, J=7.6 Hz, 1H), 6.82 (s, 1H), 5.05 (d, J=11.7 Hz, 2H), 4.56 (s , 2H), 4.52-4.46 (m, 2H), 3.09 (s, 6H), 2.92 (s, 3H), 1.88-1.76 (m, 2H), 1, 40-1.19 (m, 9H), 0.87 (t, J=7.4 Hz, 3H); LCMS m/z (ESI+) for (C ₂₆ H ₃₄ N ₈ O ₃ ), 507.4 (M+H) ⁺ .

Stage 2: Example 300

To a suspension of tert-butyl ({6-(dimethylamino)-1-oxo-2-[6-(4-propyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl]-2, 3-dihydro-1H-pyrrolo[3.4-c]pyridin-4-yl}methyl)methylcarbamate (1.96 g, 3.87 mmol) in MeOH (20 ml) add a solution of HCl (4.0 M in 1.4 -dioxane, 19.3 ml, 77.4 mmol) slowly at 0°C. The mixture is stirred for 3 hours at 0°C and then slowly warmed to room temperature. The mixture is stirred at room temperature for 16 hours. The reaction is concentrated to dryness. The solids are dissolved in 1:9 MeOH/DCM (80 ml), cooled to 0°C and then stirred with saturated Na ₂ CO ₃ solution (25 ml) for 20 minutes. The mixture is separated. The aqueous layer was extracted with a mixture of 1:19 MeOH/DCM (3×50 ml). The combined organic layers are washed with H ₂ O (2×30 ml), dried over Na ₂ SO ₄ , filtered and concentrated. The solids are suspended in EtOAc at 40°C for 40 minutes. Solids are collected by filtration. The filter cake was washed with EtOAc and then dried for 16 hours in a vacuum oven at 30°C to give Example 300 (1.42 g, 90% yield) as a yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.73 (s, 1H), 8.62 (dd, J=8.4, 1.0 Hz, 1H), 8.11-8.04 ( m, 1H), 8.00 (dd, J=1.7, 1.0 Hz, 1H), 6.78 (s, 1H), 5.14 (s, 2H), 4.57 (dd, d =7.9, 6.5 Hz, 2H), 3.80 (s, 2H), 3.09 (s, 6H), 2.35 (s, 3H), 1.88 (g, J=1, 4 Hz, 2H), 0.93 (t, J=1.4 Hz, 3H); LCMS m/z (ESI+) for (C ₂₁ H ₂₆ N ₈ O), 407.3 (M+H) ⁺ .

Biological assays and data

Biochemical enzyme assay HPK1

Inhibition of the HPK1 enzyme is measured using a fluorescence-based chelation-enhanced fluorescence (CHEF) method (1) and using a proprietary fluorescent peptide substrate in which a cysteine residue is alkylated with a sulfonamidoxine derivative to produce an amino acid called C-Sox (CSx). Reactions are carried out in 50 μl volumes in 96-well plates and contain 0.5 nM full-length autophosphorylated recombinant human HPK1 (see below for preparation method), 3 μM phosphoacceptor peptide substrate (Ac-[CSx]HSLPRFNR-amide peptide substrate, also known as AQT0178, AssayQuant Technologies Inc., Hopkinton, MA), test compound (11 doses, 3-fold serial dilutions, final concentration 2% DMSO) or DMSO alone, 0.002% Tween-20, 1 mM DTT and 2.5 mM MgCl ₂ in 50 mM MOPS (3-(N-morpholino)propanesulfonic acid) buffer, pH 7.8, and initiated by adding 45 μM ATP after a 20-minute pre-incubation. The initial reaction rates are determined by monitoring the fluorescence of the peptide (λ _ex =360 nm, λ _em =500 nm) at 30°C for 15 minutes on a Tecan M1000 plate reader (Tecan Group Ltd., Mannedorf, Zurich, Switzerland). Inhibition constant (K _i ) values are calculated by fitting the initial reaction rates to the Morrison equation (2) for competitive inhibition with strong binding using a nonlinear regression method and the experimentally measured apparent K _m of ATP (19 μM). Kinetic and crystallographic studies indicate that the inhibitors are ATP-competitive.

Generation of autophosphorylated full-length recombinant HPK1

The DNA sequence encoding full-length human HPK1 (Genbank NM_001042600.2) is synthesized in GenScript and contains a limited number of silent base substitutions to eliminate specific restriction endonuclease recognition sites, and is fused to the DNA sequence encoding tobacco etch virus (TEV) N-. terminal polyhistidine tag for purification. Recombinant baculovirus was produced using the Bac-to-Bac method (InVitrogen) and used to infect 9 L of ST21 insect cells. All purification stages are carried out at 4°C. Cells containing recombinant HPK1 were lysed in 50 mM HEPES-NaOH, pH 7.5, 250 mM NaCl, 1 mM tris(2-carboxyethyl)phosphine (TCEP), 10 μM leupeptin (Sigma), 10 μM E-64 (Sigma ) and one “EDTA-free” protease inhibitor tablet (Roche) per 75 ml buffer, centrifuged at 15,000 for 1 hour, and the supernatant passed through 5 ml ProBond IMAC resin (InVitrogen). After washing, HPK1 was eluted with 12.5 ml of 50 mM HEPES-NaOH, pH 7.5, 400 mM NaCl, 250 mM imidazole-HCl, pH 7.5, 1 mM TCEP, 2 μM leupeptin, and 2 μM E-64. Eluted HPK1 is dialyzed overnight against 0.8 L of 50 mM HEPES-NaOH, pH 7.5, 400 mM NaCl, 20 mM imidazole-HCl, pH 7.5, 1 mM TCEP, 2 mM leupeptin, 2 mM E-64 , 2 mM ATP-NaOH pH 7.3, 5 mM MgCl ₂ and 1 mg TEV (this step simultaneously reduces the imidazole concentration in preparation for the subsequent reverse IMAC chromatography step, removes the polyhistidine purification tag with TEV protease, and autophosphorylates HPK1 in the presence MdATP). Detagged and autophosphorylated HPK1 was adjusted to 40 mM imidazole-HCl, pH 7.5, and passed through a 10 mL ProBond resin column equilibrated in a similar manner. Unlabeled HPK1 passing through the column was concentrated to approximately 7 mL and further purified by gel filtration through a Superdex 26/60 size exclusion column (GE Healthcare) equilibrated and treated in 41.7 mM HEPES-NaOH buffer, pH 7.5, containing 417 mM NaCl and 0.417 mM TCER. Peak fractions containing HPK1 were pooled and concentrated to approximately 2.5 ml using a centrifugal concentrator (Millipore). Concentrated HPK1 is adjusted to 20% (v/v) glycerol by adding 50% (v/v) glycerol, resulting in a final composition of 25 mM HEPES-NaOH, pH 7.5, 250 mM NaCl, 0. 25 mM TCER and 20% (v/v) glycerol. The final protein concentration is approximately 1 mg/ml, as determined by the Bradford assay (Pearce's reagent) and using bovine serum albumin (Pearce standard 2 mg/ml) as the protein standard. Mass spectrogram analysis shows that autophosphorylated HPK1 contains an average of 15 phosphates per monomer. Aliquots of the enzyme are flash frozen in liquid nitrogen and stored at -80°C. The final product contains a glycine-serine extension at the N-terminus of the full-length protein as a result of the cloning strategy.

The final DNA sequence of full-length HPK1 with an N-terminal tag (SEQ ID NO. 1) includes the post-solidus sequence after removal of the purification tag (underlined), which encodes full-length HPK1 with an additional 5'-GGATCC BamR I cloning site sequence (SEQ ID NO. 3 ):

The final protein sequence of N-terminally tagged full-length HPK1 (SEQ ID NO. 2) includes the sequence after the solidus after removal of the purification tag (underlined) with an additional GlySer sequence at the N-terminus encoded from the BamH I cloning site (SEQ ID NO. 4) :

Cellular assays

Phospho-SLP-76 (Ser376) homogeneous time-resolved fluorescence (HTRF) assay

7urkat cells were seeded at 90,000 cells/well in 90 μl of RPMI1640 growth medium containing 10% FBS and incubated at 37°C with 5% CO ₂ overnight. The next day, compounds are serially diluted, starting at a maximum dose of 10 mM, to obtain an 11-point curve of 3-fold dilution in DMSO. Compounds were intermediately diluted 1:100 in growth medium before diluting 1:10 on cells to a final concentration of 10 μM to 0.1 nM in 0.1% DMSO. After a 30-minute pretreatment with compounds, cells were stimulated with 200 μg/ml anti-CD3 complexed with F(ab)2 (clone UCTH1) for 15 minutes at 37°C with 5% CO ₂ . Stimulation is stopped with ice-cold PBS, and cells are collected by centrifugation before lysis in Cisbio lysis buffer (Cisbio, Bedford, MA). Lysates were transferred to small volume white plates containing anti-phospho-SLP-76-cryptate plus anti-phospho-SLP-76-d2 HTRF antibodies and incubated overnight at room temperature, protected from light according to the manufacturer's protocol (Cisbio , Bedford, MA). HTRF is measured on a Perkin Elmer Envision and IC ₅₀ values are calculated by fitting a concentration-response curve using four-parameter nonlinear regression analysis.

Data on the biological activity of the compounds are presented in table 8. Data for HPK1 CHEF are presented in table 8 in the column headed HPK1 Ki (μM). Data for cell-based homogeneous time-resolved fluorescence (HTRF) assays of Phospho-SLP-76 (Ser376) are presented in Table 8 under the column heading pSLP76 IC ₅₀ (µM). For each of these two assays, the corresponding adjacent column indicates how many times the compound was tested in the assay.

Biochemical Kinase Selectivity Analysis

PRKD2 (PKD2) and RPS6KA2 (RSK3) kinase assays were performed at CT ATP levels at Thermo Fisher Scientific, Inc. (Madison, WI) using their Invitrogen™SelectScreen™ for Z'-LYTE™ fluorescence resonance energy transfer (FRET) based on the different sensitivities of phosphorylated and non-phosphorylated peptides to proteolytic cleavage. Recombinant full-length human GST-tagged PRKD2 (PKD2) and RPS6KA2 (RSK3) are produced by Thermo Fisher Scientific, Inc. (Madison, WI). Kinase assays are performed in 10-μl reaction mixtures. PRKD2 (PKD2) reactions contain 0.64-5.84 ng enzyme, 25 µM ATP (~Km), 2 µM Ser/Thr 17 (Z'-LYTE™ peptide substrate), 10 mM MgCl2, 0.01% BRIJ- 35, 1 mM EGTA in 50 mM HE PES, pH 7.5. RPS6KA2 (RSK3) reactions are carried out similarly and contain 0.5-9 ng enzyme or 10 µM ATP (~Km) and 2 µM Ser/Thr 06 peptide. After a 1-hour incubation of the kinase reaction, the PRKD2 (PKD2) and RPS6KA2 (RSK3), respectively, add 5 μl of Z'-LYTE™ Development Reagent A solution at a dilution of 1:256 and 1:4096. The extent of kinase reactions leading to a change in the FRET signal of the peptide substrate is measured, and the inhibition of each kinase is measured relative to the DMSO control and reported as the average of replicate measurements. Each 50% inhibitory concentration (IC ₅₀ ) determination is based on 10-dose replicate measurements corresponding to the standard four-parameter IC ₅₀ equation using XLfit software from IDBS (Guildford, United Kingdom). The dose-response curve corresponds to model curve number 205 (sigmoidal dose-response model). The IC ₅₀ ratio of PRKD2 to HPK1 Ki and the IC ₅₀ ratio of RSK3 to HPK1 Ki are presented in the corresponding columns adjacent to those for PRKD2 IC ₅₀ and RSK3 IC ₅₀ .

Alpha branched amine analogues, where at least one of R ^3a and R ^3b is other than hydrogen, significantly improve selectivity for HPK1 over other kinases (including PRKD2 and RSK3) compared to non-alpha branched amine compounds , for example, Example 300. Cyclic, fused triazoles, for example, compounds of formula IB, where R ⁴ is (R ⁴ -ii), provide improved biochemical and cellular effectiveness compared to non-cyclic compounds, for example, Example 300.

An empty field in Table 8 means that no data was received.

All publications and patent applications cited in the specification are incorporated herein by reference in their entirety. It will be apparent to those skilled in the art that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

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LIST OF SEQUENCES

<110> Pfizer Inc.

Barber, Joyann

Cho-Schultz, Sujin

Del Bel, Matthew L.

Gallego, Rebecca Anne

He, Mingying

Jalaie, Mehran

Kania, Robert Steven

McTigue, Michele Ann

Nair, Sajiv Krishnan

Schmitt, Anne-Marie Dechert

Tuttle, Jamison Bryce

Zhou, Dahui

Zhou, Ru

<120> Azalactam compounds as HPK1 inhibitors

<130>PC72605A

<150>US 63/018,689

<151> 2020-05-01

<160> 4

<170> Patent In version 3.5

<210> 1

<211> 2472

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tcaggggacc tggtggcact gaagatggtg aagatggagc ctgatgatga tgtctccacc 180

cttcagaagg aaatcctcat attgaaaact tgccggcacg ccaacatcgt ggcctaccac 240

gggagttatc tctggttgca gaaactctgg atctgcatgg agttctgtgg ggctggttct 300

ctccaggaca tctaccaagt gacaggctcc ctgtcagagc tccagattag ctatgtctgc 360

cgggaagtgc tccagggact ggcctatttg cactcacaga agaagataca cagggacatc 420

aagggagcta acatcctcat caatgatgct ggggaggtca gattggctga ctttggcatc 480

tcggcccaga ttggggctac actggccaga cgcctctctt tcattgggac accctactgg 540

atggctccgg aagtggcagc tgtggccctg aagggaggat acaatgagct gtgtgacatc 600

tggtccctgg gcatcacggc catcgaactg gccgagctac agccaccgct ctttgatgtg 660

caccctctca gagttctctt cctcatgacc aagagtggct accagcctcc ccgactgaag 720

gaaaaaggca aatggtcggc tgccttccac aacttcatca aagtcactct gactaagagt 780

cccaagaaac gacccagcgc caccaagatg ctcagtcatc aactggtatc ccagcctggg 840

ctgaatcgag gcctgatcct ggatcttctt gacaaactga agaatcccgg gaaaggaccc 900

tccattgggg acattgagga tgaggagccc gagctacccc ctgctatccc tcggcggatc 960

agatccaccc accgctccag ctctctgggc atcccagatg cagactgctg tcggcggcac 1020

atggagttca ggaagctccg aggaatggag accagacccc cagccaacac cgctcgccta 1080

cagcctcccc gagacctcag gagcagcagc cccaggaagc aactgtcaga gtcgtctgac 1140

gatgactatg acgacgtgga catccccacc cctgcagagg acacacctcc tccacttccc 1200

cccaagccca agttccgttc tccatcagac gagggtcctg ggagcatggg ggatgatggg 1260

cagctgagcc cgggggtgct ggtccggtgt gccagtgggc ccccaccaaa cagcccccgt 1320

cctgggcctc ccccatccac cagcagcccc cacctcaccg cccattcaga accctcactc 1380

tggaacccac cctcccggga gcttgacaag cccccacttc tgccccccaa gaaggaaaag 1440

atgaagagaa agggatgtgc ccttctcgta aagttgttca atggctgccc cctccgcatc 1500

cacagcacgg ccgcctggac acatccctcc accaaggacc agcacctgct cctgggggca 1560

gaggaaggca tcttcatcct gaaccggaat gaccaggagg ccacgctgga aatgctcttt 1620

cctagccgga ctacgtgggt gtactccatc aacaacgttc tcatgtctct ctcaggaaag 1680

accccccacc tgtattctca tagcatcctt ggcctgctgg aacggaaaga gaccagagca 1740

ggaaacccca tcgctcacat tagcccccac cgcctactgg caaggaagaa catggtttcc 1800

accaagatcc aggacaccaa aggctgccgg gcgtgctgtg tggcggaggg tgcgagctct 1860

gggggcccgt tcctgtgcgg tgcattggag acgtccgttg tcctgcttca gtggtaccag 1920

cccatgaaca aattcctgct tgtccggcag gtgctgttcc cactgccgac gcctctgtcc 1980

gtgttcgcgc tgctgaccgg gccaggctct gagctgcccg ctgtgtgcat cggcgtgagc 2040

cccggggcggc cggggaagtc ggtgctcttc cacacggtgc gctttggcgc gctctcttgc 2100

tggctgggcg agatgagcac cgagcacagg ggacccgtgc aggtgaccca ggtagaggaa 2160

gatatggtga tggtgttgat ggatggctct gtgaagctgg tgaccccgga ggggtcccca 2220

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gatcagctgc tacaggagct gagagaccct accctcactt tccgtctgct tggctccccc 2400

aggcctgtag tggtggagac acgcccagtg gatgatccta ctgctcccag caacctctac 2460

atccaggaatga 2472

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Asp His Tyr Asp Leu Leu Gln Arg Leu Gly Gly Gly Thr Tyr Gly Glu

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Val Phe Lys Ala Arg Asp Lys Val Ser Gly Asp Leu Val Ala Leu Lys

35 40 45

Met Val Lys Met Glu Pro Asp Asp Asp Val Ser Thr Leu Gln Lys Glu

50 55 60

Ile Leu Ile Leu Lys Thr Cys Arg His Ala Asn Ile Val Ala Tyr His

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Gly Ser Tyr Leu Trp Leu Gln Lys Leu Trp Ile Cys Met Glu Phe Cys

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Gly Ala Gly Ser Leu Gln Asp Ile Tyr Gln Val Thr Gly Ser Leu Ser

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Glu Leu Gln Ile Ser Tyr Val Cys Arg Glu Val Leu Gln Gly Leu Ala

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Tyr Leu His Ser Gln Lys Lys Ile His Arg Asp Ile Lys Gly Ala Asn

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Ile Leu Ile Asn Asp Ala Gly Glu Val Arg Leu Ala Asp Phe Gly Ile

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Ser Ala Gln Ile Gly Ala Thr Leu Ala Arg Arg Leu Ser Phe Ile Gly

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Thr Pro Tyr Trp Met Ala Pro Glu Val Ala Ala Val Ala Leu Lys Gly

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Gly Tyr Asn Glu Leu Cys Asp Ile Trp Ser Leu Gly Ile Thr Ala Ile

195 200 205

Glu Leu Ala Glu Leu Gln Pro Pro Leu Phe Asp Val His Pro Leu Arg

210 215 220

Val Leu Phe Leu Met Thr Lys Ser Gly Tyr Gln Pro Pro Arg Leu Lys

225 230 235 240

Glu Lys Gly Lys Trp Ser Ala Ala Phe His Asn Phe Ile Lys Val Thr

245 250 255

Leu Thr Lys Ser Pro Lys Lys Arg Pro Ser Ala Thr Lys Met Leu Ser

260 265 270

His Gln Leu Val Ser Gln Pro Gly Leu Asn Arg Gly Leu Ile Leu Asp

275 280 285

Leu Leu Asp Lys Leu Lys Asn Pro Gly Lys Gly Pro Ser Ile Gly Asp

290 295 300

Ile Glu Asp Glu Glu Pro Glu Leu Pro Pro Ala Ile Pro Arg Arg Ile

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Arg Ser Thr His Arg Ser Ser Ser Leu Gly Ile Pro Asp Ala Asp Cys

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Cys Arg Arg His Met Glu Phe Arg Lys Leu Arg Gly Met Glu Thr Arg

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Pro Pro Ala Asn Thr Ala Arg Leu Gln Pro Pro Arg Asp Leu Arg Ser

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Ser Ser Pro Arg Lys Gln Leu Ser Glu Ser Ser Asp Asp Asp Tyr Asp

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Asp Val Asp Ile Pro Thr Pro Ala Glu Asp Thr Pro Pro Pro Leu Pro

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Pro Lys Pro Lys Phe Arg Ser Pro Ser Asp Glu Gly Pro Gly Ser Met

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Gly Asp Asp Gly Gln Leu Ser Pro Gly Val Leu Val Arg Cys Ala Ser

420 425 430

Gly Pro Pro Pro Asn Ser Pro Arg Pro Gly Pro Pro Pro Ser Thr Ser

435 440 445

Ser Pro His Leu Thr Ala His Ser Glu Pro Ser Leu Trp Asn Pro Pro

450 455 460

Ser Arg Glu Leu Asp Lys Pro Pro Leu Leu Pro Pro Lys Lys Glu Lys

465 470 475 480

Met Lys Arg Lys Gly Cys Ala Leu Leu Val Lys Leu Phe Asn Gly Cys

485 490 495

Pro Leu Arg Ile His Ser Thr Ala Ala Trp Thr His Pro Ser Thr Lys

500 505 510

Asp Gln His Leu Leu Leu Gly Ala Glu Glu Gly Ile Phe Ile Leu Asn

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Arg Asn Asp Gln Glu Ala Thr Leu Glu Met Leu Phe Pro Ser Arg Thr

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Thr Trp Val Tyr Ser Ile Asn Asn Val Leu Met Ser Leu Ser Gly Lys

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Thr Pro His Leu Tyr Ser His Ser Ile Leu Gly Leu Leu Glu Arg Lys

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Glu Thr Arg Ala Gly Asn Pro Ile Ala His Ile Ser Pro His Arg Leu

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Leu Ala Arg Lys Asn Met Val Ser Thr Lys Ile Gln Asp Thr Lys Gly

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Cys Arg Ala Cys Cys Val Ala Glu Gly Ala Ser Ser Gly Gly Pro Phe

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Leu Cys Gly Ala Leu Glu Thr Ser Val Val Leu Leu Gln Trp Tyr Gln

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Pro Met Asn Lys Phe Leu Leu Val Arg Gln Val Leu Phe Pro Leu Pro

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Thr Pro Leu Ser Val Phe Ala Leu Leu Thr Gly Pro Gly Ser Glu Leu

660 665 670

Pro Ala Val Cys Ile Gly Val Ser Pro Gly Arg Pro Gly Lys Ser Val

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Leu Phe His Thr Val Arg Phe Gly Ala Leu Ser Cys Trp Leu Gly Glu

690 695 700

Met Ser Thr Glu His Arg Gly Pro Val Gln Val Thr Gln Val Glu Glu

705 710 715 720

Asp Met Val Met Val Leu Met Asp Gly Ser Val Lys Leu Val Thr Pro

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Glu Gly Ser Pro Val Arg Gly Leu Arg Thr Pro Glu Ile Pro Met Thr

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Glu Ala Val Glu Ala Val Ala Met Val Gly Gly Gln Leu Gln Ala Phe

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Trp Lys His Gly Val Gln Val Trp Ala Leu Gly Ser Asp Gln Leu Leu

770 775 780

Gln Glu Leu Arg Asp Pro Thr Leu Thr Phe Arg Leu Leu Gly Ser Pro

785 790 795 800

Arg Pro Val Val Val Glu Thr Arg Pro Val Asp Asp Pro Thr Ala Pro

805 810 815

Ser Asn Leu Tyr Ile Gln Glu

820

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<213> Homo sapiens

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tttcag 66

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Glu Asn Leu Tyr Phe Gln

20

<---

Claims (67)

1. Compound of formula I: or a pharmaceutically acceptable salt or stereoisomer thereof, wherein: R ¹ represents (C ₁ -C ₆ )alkyl, -N(R ⁵ )(R ⁶ ), or (C ₃ -C ₆ )cycloalkyl, wherein said (C ₃ -C ₆ )cycloalkyl is substituted with 0 or 1 substituent, which is (C ₁ -C ₆ )alkyl or (C ₁ -C ₆ )alkoxy, where: R ⁵ and R ⁶ are each independently hydrogen or (C ₁ -C ₆ )alkyl, or R ⁵ and R ⁶ together with the nitrogen to which they are attached form a (4-8 membered) heterocycloalkyl which is substituted with 0, 1 or 2 substituents independently selected from the group consisting of (C ₁ -C ₆ )alkyl, halogen (C ₁ -C ₆ )alkyl and (C ₁ -C ₆ )alkoxy; R ² represents N(R ⁷ )(R ⁸ ), where: each of R ⁷ and R ⁸ independently represents hydrogen or (C ₁ -C ₆ )alkyl, which is substituted with 0 or 1 substituent representing halogen, (C ₁ -C ₆ )alkoxy, cyano or hydroxy; R ^3a represents hydrogen or (C ₁ -C ₃ )alkyl substituted with 0 or 1 hydroxy or (C ₁ -C ₃ )alkoxy substituent; R ^3b is hydrogen or (C ₁ -C ₃ )alkyl, provided that R ^3a and R ^3b are both not H when R ⁴ is (R ⁴ -i); R ⁴ is (R ⁴ -i) or (R ⁴ -ii): Where: R ^4N is (C ₁ -C ₆ )alkyl, halogen(C ₁ -C ₆ )alkyl or (C ₃ -C ₆ )cycloalkyl; R ^4C is hydrogen, (C ₁ -C ₆ )alkyl, halogen(C ₁ -C ₆ )alkyl or (C ₁ -C ₆ )alkoxy; R ^4D represents hydrogen, (C ₁ -C ₆ )alkyl, halogen(C ₁ -C ₆ )alkyl, halogen(C ₁ -C ₆ )alkoxy or (C ₃ -C ₆ )cycloalkyl, wherein said (C ₁ - C ₆ )alkyl and halogen(C ₁ -C ₆ )alkyl are substituted with 0 or 1 substituent, which is hydroxy or (C ₁ -C ₆ )alkoxy; R ^4E represents hydrogen or (C ₁ -C ₆ )alkyl; And R ^4F represents hydrogen, halogen, (C ₁ -C ₆ )alkyl or halogen(C ₁ -C ₆ )alkyl, wherein said (C ₁ -C ₆ )alkyl and halogen(C ₁ -C ₆ )alkyl are substituted with 0 or 1 substituent, which is hydroxy, cyano or (C ₁ -C ₆ )alkoxy. 2. A compound according to claim 1 or a pharmaceutically acceptable salt or stereoisomer thereof, characterized in that R ⁴ is (R ⁴ -i); R ^4N represents (C ₁ -C ₆ )alkyl or (C ₃ -C ₆ )cycloalkyl; And R ^4C represents hydrogen or (C ₁ -C ₃ )alkyl. 3. A compound according to claim 1 or 2 or a pharmaceutically acceptable salt or stereoisomer thereof, characterized in that R ¹ represents -N(R ⁵ )(R ⁶ ) or (C ₃ -C ₆ )cycloalkyl, wherein said (C ₃ -C ₆ )cycloalkyl is cyclopropyl and is substituted with 0 or 1 substituent which is (C ₁ -C ₆ )alkyl, wherein said (C ₁ -C ₆ )alkyl is methyl; R ⁵ and R ⁶ are each independently hydrogen or (C ₁ -C ₃ )alkyl, or R ⁵ and R ⁶ together with the nitrogen atom to which they are attached form a (5-membered) heterocycloalkyl substituted with 0 or 1 substituent, which is (C ₁ -C ₆ )alkyl; R ² is N(R ⁷ )(R ⁸ ), wherein R ⁷ and R ⁸ are each independently hydrogen or (C ₁ -C ₆ )alkyl, which is methyl; R ^3a is (C ₁ -C ₃ )alkyl substituted with 0 or 1 substituent, which is (C ₁ -C ₃ )alkoxy; And R ^3b represents hydrogen or (C ₁ -C ₃ )alkyl. 4. The compound according to claim 1 or a pharmaceutically acceptable salt or stereoisomer thereof, characterized in that R ⁴ represents (R ⁴ -ii); R ^4D represents hydrogen, (C ₁ -C ₆ )alkyl or halogen(C ₁ -C ₆ )alkyl, wherein said (C ₁ -C ₆ )alkyl is substituted with 0 or 1 substituent which is hydroxy; R ^4E represents hydrogen; And R ^4F represents hydrogen, (C ₁ -C ₆ )alkyl or halogen(C ₁ -C ₆ )alkyl, wherein said (C ₁ -C ₆ )alkyl is substituted with 0 or 1 substituent which is hydroxy. 5. A compound according to claim 1 or 4 or a pharmaceutically acceptable salt or stereoisomer thereof, characterized in that R ¹ represents -N(R ⁵ )(R ⁶ ) or (C ₃ -C ₆ )cycloalkyl, wherein said (C ₃ -C ₆ )cycloalkyl is cyclopropyl and is substituted with 0 or 1 substituent which is (C ₁ -C ₆ )alkyl, wherein said (C ₁ -C ₆ )alkyl is methyl; R ⁵ and R ⁶ are each independently hydrogen or (C ₁ -C ₃ )alkyl, or R ⁵ and R ⁶ together with the nitrogen atom to which they are attached form a (5-membered) heterocycloalkyl substituted with 0 or 1 substituent, which is (C ₁ -C ₆ )alkyl; R ² represents N(R ⁷ )(R ⁸ ), wherein each of R ⁷ and R ⁸ independently represents hydrogen or (C ₁ -C ₆ )alkyl, which represents methyl; R ^3a represents hydrogen or (C ₁ -C ₃ )alkyl substituted with 0 or 1 substituent, which is (C ₁ -C ₃ )alkoxy; And R ^3b represents hydrogen or (C ₁ -C ₃ )alkyl. 6. Connection according to any one of paragraphs. 1-5 or a pharmaceutically acceptable salt or stereoisomer thereof, characterized in that R ¹ is 2-methylpyrrolidin-1-yl or 2(R)-methylpyrrolidin-1-yl. 7. Connection according to any one of paragraphs. 1-6 or a pharmaceutically acceptable salt or stereoisomer thereof, characterized in that R ⁷ and R ⁸ are each hydrogen. 8. Connection according to any one of paragraphs. 1-6 or a pharmaceutically acceptable salt or stereoisomer thereof, characterized in that R ⁷ represents hydrogen; and R ⁸ represents (C ₁ -C ₃ )alkyl. 9. Connection according to any one of paragraphs. 1-8 or a pharmaceutically acceptable salt or stereoisomer thereof, characterized in that R ^3b is hydrogen, and wherein the orientation of R ² , R ^3a and R ^3b gives a compound of formula I(R) or formula I(S): 10. Connection according to any one of paragraphs. 1 and 4-9 or a pharmaceutically acceptable salt or stereoisomer thereof, characterized in that R ⁴ is 5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol- 3-yl, (5R)-5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl, (5S)-5-methyl-6 ,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl, 5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1 ,2,4]triazol-3-yl, (5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl or (5R )-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl, wherein said methyl is substituted with 0 or 1 substituent which is F or OH . 11. The compound according to claim 10 or a pharmaceutically acceptable salt or stereoisomer thereof, characterized in that R ⁴ is (5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1, 2,4]triazol-3-yl or (5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl, where the indicated is methyl substituted with 0 or 1 substituent which is F or OH. 12. The compound representing 4-[1-aminopropyl]-2-{6-[5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole-3 -yl]pyridin-2-yl}-6-[2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one; 4-[1-aminoethyl]-2-{6-[5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl]pyridin-2 -yl}-6-[2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one; 4-[1-aminoethyl]-2-{6-[5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl]pyridin-2 -yl}-6-[2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one; 4-[1-aminoethyl]-2-{6-[5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-e][1,2,4]triazol-3-yl]pyridin-2 -yl}-6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one; 4-[1-aminopropyl]-2-{3-[5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazol-3-yl]phenyl}- 6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one; or 4-[1-amino-2-methoxyethyl]-6-(1-methylcyclopropyl)-2-{6-[5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2 ,4]triazol-3-yl]pyridin-2-yl}-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one. 13. The compound according to claim 12, characterized in that the compound is 4-[(1R)-1-aminopropyl]-2-{6-[(5S)-5-methyl-6,7-dihydro-5H-pyrrolo [2,1-c][1,2,4]triazol-3-yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H -pyrrolo[3,4-c]pyridin-1-one; 4-[(1R)-1-aminoethyl]-2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole -3-yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one; 4-[(1S)-1-aminoethyl]-2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole -3-yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one; 4-[(1R)-1-aminoethyl]-2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole -3-yl]pyridin-2-yl}-6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one; 4-[(1R)-1-aminopropyl]-2-{3-[(5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazole -3-yl]phenyl}-6-[methyl(propan-2-yl)amino]-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one; 4-[(1R)-1-amino-2-methoxyethyl]-6-(1-methylcyclopropyl)-2-{6-[(5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2, 1-c][1,2,4]triazol-3-yl]pyridin-2-yl}-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one; or 4-[(1S)-1-amino-2-methoxyethyl]-6-(1-methylcyclopropyl)-2-{6-[(5S)-5-methyl-6,7-dihydro-5H-pyrrolo[2, 1-c][1,2,4]triazol-3-yl]pyridin-2-yl}-2,3-dihydro-1H-pyrrolo[3,4-c]pyridin-1-one. 14. A compound having the structure: 15. The compound representing 4-[(1R)-1-aminoethyl]-2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1 ,2,4]triazol-3-yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c] pyridin-1-one. 16. The compound representing 4-[(1S)-1-aminoethyl]-2-{6-[(5S)-5-ethyl-6,7-dihydro-5H-pyrrolo[2,1-c][1 ,2,4]triazol-3-yl]pyridin-2-yl}-6-[(2R)-2-methylpyrrolidin-1-yl]-2,3-dihydro-1H-pyrrolo[3,4-c] pyridin-1-one. 17. Pharmaceutical composition for inhibiting HPK1, containing a therapeutically effective amount of a compound according to any one of paragraphs. 1-16 or a pharmaceutically acceptable salt or stereoisomer thereof and a pharmaceutically acceptable carrier or excipient. 18. A method of treating abnormal cell growth associated with inhibition of HPK1 activity in a mammal, comprising administering to the mammal a therapeutically effective amount of a compound according to any one of claims. 1-16 or a pharmaceutically acceptable salt or stereoisomer thereof. 19. The method according to claim 18, characterized in that the abnormal cell growth represents cancer. 20. A method of treating abnormal cell growth associated with inhibition of HPK1 activity in a mammal, comprising administering to the mammal a therapeutically effective amount of a compound according to any one of claims. 1-16 or a pharmaceutically acceptable salt or stereoisomer thereof, and further comprising administering an amount of an additional anti-cancer therapeutic agent, amounts of which together are effective in treating said cancer. 21. The method according to claim 20, where the cancer is selected from the group consisting of breast cancer, ovarian cancer, bladder cancer, uterine cancer, prostate cancer, lung cancer, esophageal cancer, head and neck cancer, colorectal cancer, kidney cancer , liver cancer, pancreatic cancer, stomach cancer and thyroid cancer.