Inhibitor compounds

Abstract

The present invention relates to certain compounds that function as inhibitors of PMS2 protein activity. In particular, the compounds of the present invention may be used as covalent binders to inhibit PMS2. Thus, the compounds of the present invention may be used to treat diseases or conditions mediated, at least in part, by inappropriate PMS2 activity, for example, cancer. The present invention furthermore relates to the use of the compounds and pharmaceutical compositions comprising them.

Description

INHIBITOR COMPOUNDSINTRODUCTION

[0001] The present invention relates to certain compounds that function as inhibitors of PMS2 protein activity. In particular, the compounds of the present invention may be used as covalent binders to inhibit PMS2. Thus, the compounds of the present invention may be used to treat diseases or conditions mediated, at least in part, by inappropriate PMS2 activity, for example, cancer. The present invention furthermore relates to the use of the compounds and pharmaceutical compositions comprising them.BACKGROUND OF THE INVENTION

[0002] Cancer is caused by altered cellular proliferation. Precisely what causes a cell to become malignant and proliferate in an uncontrolled and unregulated manner has been the focus of intense research over recent decades. This research has led to the identification of molecular targets associated with key pathways that enable such malignancies.

[0003] Mismatch repair (MMR) is a highly conserved DNA repair pathway that plays a major role during DNA replication, repair, and recombination, as well as during meiosis in eukaryotes and immunoglobulin maturation / diversification in mammals. MMR promotes genome stability in all organisms by correcting DNA base mismatches and insertion / deletion (indel) loops that can occasionally arise during normal DNA replication process. Base pair mismatches occur when incorrect nucleotides are inserted into the newly synthesized DNA strand and escape the proofreading function of DNA polymerases. Indel loops commonly arise in the context of microsatellites - highly polymorphic short repetitive DNA sequences distributed throughout both prokaryotic and eukaryotic genomes. Typically, at microsatellites, the template and primer strands are prone to slippage (dissociation and reannealing) during replication, which can generate loop structures and a discordant number of repeat units between the template and newly synthesized strand.

[0004] DNA mismatch repair is a bidirectional excision and re-synthesis system that initiates at a defined strand scission 3'- or 5'- to a mismatch; the excision tract extends just past the mismatch. MMR can be divided into four steps: 1) mismatch recognition by MSH proteins; 2) recruitment of MLH / PMS proteins that connect the mismatch recognition signal to where the distant DNA strand scission begins; 3) excision of the errant DNA strand; and 4) re-synthesis of the excision gap using the remaining DNA strand as a template [1], MMR is a highly conserved biological pathway. In humans, mismatch recognition by hMutSa (MSH2-MSH6) or hMutSp (MSH2-MSH3) initiates the MMR pathway. Binding of hMutSa or hMutSp to the mismatch site results in the recruitment of MutLa (MLH1-PMS2) to form a ternary complex whose proteinprotein, protein-DNA interactions and endonuclease activity are modulated by ATP / ADP cofactors. Proliferating cell nuclear antigen (PCNA) may play a role in the recruitment of MMR proteins to the vicinity of the replication fork [1], PCNA may also activate a latent endonuclease activity in eukaryotic MutLa proteins. After DNA incision, exonuclease 1 (EXO1) is recruited which excises the newly synthesized DNA strand and the DNA excision gap is re-synthesized by DNA polymerase. When DNA re-synthesis is complete, the remaining nick is ligated by DNA ligase to restore the integrity of the duplex [2], Consistent with this function, MMR is an important tumour suppressor pathway that is lost in up to 40% of sporadic cancers. Moreover, individuals with germline mutations in MMR genes develop cancer predisposition conditions.

[0005] Lynch Syndrome (LS, formerly designated as hereditary non-polyposis colorectal cancer) is the most common cause of hereditary colorectal cancer (CRC), accounting for 2-5% of all cases. LS is also characterized by an increased risk of malignancies at certain extracolonic sites such as the endometrium, ovary, stomach and small bowel, among others [3], LS has an autosomal dominant inheritance pattern and is caused by germline mutations in MMR genes MLH1 , MSH2, MSH6 or PMS2. Gene expression from the one wild-type allele is sufficient for adequate MMR activity until a second hit inactivates the wild-type allele leading to MMR deficiency.

[0006] Constitutional mismatch repair deficiency (CMMRD) syndrome is a distinct childhood cancer predisposition syndrome that results from biallelic germline mutations in one of four MMR genes, MLH1 , MSH2, MSH6 or PMS2. Patients may have either homozygous biallelic alterations or heterozygous alterations of MMR genes.

[0007] MMR-deficient cancers are commonly and typically characterized by the accumulation of DNA mutations at higher rates than normal cells and other tumours; for example, CMMRD tumours commonly have an ultra-hypermutated phenotype (>250 substitution mutations / Mb) [4], MMR deficiency also results in gains or losses in the repeat length of microsatellites, referred to as microsatellite instability (MSI). Cancers that possess more than 40% microsatellite variations (positive fortwo or more of five microsatellite markers routinely tested) are described as high frequency MSI (MSI-H). Tumours that have no MSI are microsatellite stable (MSS) and those that possess less than 40% microsatellite variations (one out of the five markers showing microsatellite instability) are low frequency MSI (MSI-L) [5], MSI analysis is a widely used diagnostic biomarker of MMR-deficient tumours and MSI status is linked with a high prevalence of frameshift (FS) mutations that can occur because of insertion / deletion within coding microsatellites. In addition to altering downstream functions of the protein, the FS creates a new amino acid sequence that serves as a substrate for antigen processing and presentation [6], stimulating the activation of CD8+ T cells (class I) and the “helper” function of CD4+ T cells (class II).

[0008] Cancers with a greater number of neoantigens are more prone to immune surveillance and have an increased likelihood of responding to immunotherapy [7]; higher neoantigen load is associated with overall lymphocytic infiltration, TILs, memory T cells, and survival in colorectal cancer [8, 9], This feature supports a rationale for immunotherapy-based treatment strategies [6], Consistent with this notion, immune checkpoint inhibitors now offer a significant therapeutic advance in the treatment of MMR-deficient (MMR- d) cancers. Inhibitors of PD-1 ; for example, pembrolizumab (Keytruda) and nivolumab (Opdivo), have been approved by the Food and Drug Administration (FDA) for patients with MMR-d or MSI-H metastatic CRC based upon the significant survival benefit they provide. The CTLA-4 inhibitor ipilimumab (Yervoy), has been approved for use in combination with nivolumab for the treatment of MMR-d or MSI-H CRC patients who were previously treated with chemotherapy. Importantly, the FDA has approved the use of pembrolizumab in MMR-d / MSI-H cancers regardless of histological tumour type

[0010] ,

[0009] It is now accepted that clinical responses to immune checkpoint inhibitors require the existence of tumour neoantigens and infiltration of T cells that recognize such neoantigens. Higher neoantigen load is associated with response to CTLA-4 and PD-1 blockade in patients with melanoma and non-small-cell lung cancer [11 , 12, 13], The number of neoantigens is linked to tumour mutational burden (TMB), and several large studies have confirmed that high TMB correlates with enhanced checkpoint inhibitor responses andimproved overall survival in certain tumour types, such as urothelial carcinoma

[0014] , non-small cell lung cancer [15-18] and small cell lung cancer

[0019] ,

[0010] Germano et al. recently proposed that MMR inactivation through silencing of MLH1 increases TMB and leads to “dynamic mutational profiles”, resulting in persistent renewal of neoantigens both in vitro and in vivo. This triggers immune surveillance and leads to the control of tumour growth, particularly in combination with immune checkpoint inhibition, in mouse models

[0020] , Similar results are observed upon silencing of MSH2

[0021] ,

[0011] Guan et al. and Lu et al. report that MLH1 deficiency leads to cytosolic DNA release, activation of the cGAS-STING pathway and IFN production. Guan et al. demonstrate that MLH1 loss leads to DNA hyperexcision, RPA exhaustion, chromosomal instability and accumulation of cytosolic DNA

[0022] , Lu et al. report that the sensing of cytosolic DNA by the cGAS STING pathway contributes to the clinical benefit of immunotherapy in patients harbouring MMR-d tumours

[0023] , Together these reports suggest that abrogation of MMR activity may elicit beneficial immune activation through activation of the cGAS-STING pathway.

[0012] MLH1 and PMS2 commonly form a heterodimer; loss of MLH1 protein typically leads to concomitant loss of PMS2 protein suggesting that either or both proteins may be essential for MMR function and cGAS / STING pathway modulation.

[0013] There is therefore a biological and clinical rationale highlighting the need for inhibitors that target the PMS2 protein, a key component of DNA MMR, to reawaken an anti-tumour immune response.

[0014] Thus, the present invention provides methods for the treatment of cancer by binding to and modulating the function of the DNA MMR component PMS2 using small molecules as single agents and in combination with immunotherapy agents, other DNA damage response pathway modulators and / or standard-of-care chemotherapeutic agents.

[0015] Outside of the cancer field, triplet repeat disorders comprise over 30 human neurodegenerative and neuromuscular inherited diseases such as Huntington’s disease (HD), myotonic dystrophy type 1 (DM1), fragile X syndrome type A (FRAXA), Friedreich’s ataxia (FRDA), and spinocerebellar ataxias (SCAs). Such disorders are characterized by the expansion of simple repeats in genomic DNA. These unstable repeats are commonly found at different regions of several genes and their expansion can cause disease by a variety of both loss- and gain-of-function pathways, for instance through interfering with the expression or properties of the gene products, or by affecting splicing or antisense regulation. Several mechanisms including errors during DNA replication, meiotic recombination, transcription, DNA repair, and chromatin remodelling have been proposed to contribute to repeat instability, which can occur at various stages of the cell cycle. There is evidence that a functional MMR pathway is required for maintaining the stability of microsatellite sequences: for example, Msh2- / - transgenic mice bearing a copy of the human HD exon 1 (containing the CAG repeats) showed reduced expansion of the introduced (CAG)n repeats when compared with Msh2+ / + HD exon 1 mice counterparts

[0024] ,

[0016] Thus, there is a further need for compounds that target components of the DNA MMR process, including PMS2, for treating triplet repeat disorders. The present invention was devised with the foregoing in mind.SUMMARY OF THE INVENTION

[0017] According to a first aspect of the present invention there is provided a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein.

[0018] According to a further aspect of the present invention, there is provided a pharmaceutical composition comprising a compound as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, in admixture with a pharmaceutically acceptable diluent or carrier.

[0019] According to a further aspect of the present invention, there is provided a pharmaceutical composition comprising a compound as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable excipient or carrier.

[0020] According to a further aspect of the present invention, there is provided a method of inhibiting PMS2 activity, in vitro or in vivo, said method comprising contacting a cell with a therapeutically effective amount of a compound or a pharmaceutically acceptable salt, hydrate or solvate thereof as defined herein, or a pharmaceutical composition as defined herein.

[0021] According to a further aspect of the present invention, there is provided a method of treating a disease or disorder in which PMS2 activity is implicated in a patient in need of such treatment, said method comprising administering to said patient a therapeutically effective amount of a compound or a pharmaceutically acceptable salt, hydrate or solvate thereof as defined herein, or a pharmaceutical composition as defined herein.

[0022] According to a further aspect of the present invention, there is provided a method of treating a proliferative disorder in a patient in need of such treatment, said method comprising administering to said patient a therapeutically effective amount of a compound or a pharmaceutically acceptable salt, hydrate or solvate thereof as defined herein, or a pharmaceutical composition as defined herein.

[0023] According to a further aspect of the present invention, there is provided a method of treating cancer in a patient in need of such treatment, said method comprising administering to said patient a therapeutically effective amount of a compound or a pharmaceutically acceptable salt, hydrate or solvate thereof as defined herein, or a pharmaceutical composition as defined herein. In a particular embodiment, the cancer is human cancer.

[0024] According to a further aspect of the present invention, there is provided a method of treating a triplet repeat disorder in a patient in need of such treatment, said method comprising administering to said patient a therapeutically effective amount of a compound or a pharmaceutically acceptable salt, hydrate or solvate thereof as defined herein, or a pharmaceutical composition as defined herein. In a particular embodiment, the triplet repeat disorder is selected from the group consisting of Huntington’s disease (HD), myotonic dystrophy type 1 (DM1), fragile X syndrome type A (FRAXA), Friedreich’s ataxia (FRDA), and spinocerebellar ataxias (SCAs).

[0025] According to a further aspect of the present invention, there is provided a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein for use in therapy.

[0026] According to a further aspect of the present invention, there is provided a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein for use as a medicament.

[0027] According to a further aspect of the present invention, there is provided a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein, or a pharmaceutical composition as defined herein for use in the inhibition of PMS2 activity.

[0028] According to a further aspect of the present invention, there is provided a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein, or a pharmaceutical composition as defined herein for use in the treatment of a disease or disorder in which PMS2 activity is implicated.

[0029] According to a further aspect of the present invention, there is provided a compound or a pharmaceutically acceptable salt, hydrate or solvate thereof as defined herein, or a pharmaceutical composition as defined herein, for use in the treatment of a proliferative disorder.

[0030] According to a further aspect of the present invention, there is provided a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein for use in the treatment of cancer. In a particular embodiment, the cancer is human cancer.

[0031] According to a further aspect of the present invention, there is provided a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein for use in the treatment of a triplet repeat disorder. In a particular embodiment, the triplet repeat disorder is selected from the group consisting of Huntington’s disease (HD), myotonic dystrophy type 1 (DM1), fragile X syndrome type A (FRAXA), Friedreich’s ataxia (FRDA), and spinocerebellar ataxias (SCAs).

[0032] According to a further aspect of the present invention, there is provided a use of a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein, or a pharmaceutical composition as defined herein in the manufacture of a medicament for the inhibition of PMS2 activity.

[0033] According to a further aspect of the present invention, there is provided a use of a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein, or a pharmaceutical composition as defined herein in the manufacture of a medicament for the treatment of a disease or disorder in which PMS2 activity is implicated.

[0034] According to a further aspect of the present invention, there is provided the use of a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein, or a pharmaceutical composition as defined herein in the manufacture of a medicament for the treatment of a proliferative disorder.

[0035] According to a further aspect of the present invention, there is provided the use of a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein, or a pharmaceutical composition as defined herein in the manufacture of a medicament for the treatment of cancer. In a particular embodiment, the cancer is human cancer.

[0036] According to a further aspect of the present invention, there is provided the use of a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein, or a pharmaceutical composition as defined herein in the manufacture of a medicament for the treatment of a triplet repeat disorder. In a particular embodiment, the triplet repeat disorder is selected from the group consisting of Huntington’s disease (HD), myotonic dystrophy type 1 (DM1), fragile X syndrome type A (FRAXA), Friedreich’s ataxia (FRDA), and spinocerebellar ataxias (SCAs).

[0037] According to a further aspect of the present invention, there is provided a process for preparing a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein.

[0038] According to a further aspect of the present invention, there is provided a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, obtainable by, or obtained by, or directly obtained by a process of preparing a compound as defined herein.

[0039] According to a further aspect of the present invention, there are provided novel intermediates as defined herein which are suitable for use in any one of the synthetic methods set out herein.

[0040] In the above-outlined aspects of the invention, the proliferative disorder is suitably cancer, and the cancer is suitably a human cancer. In particular, the compounds and compositions of the present invention will be useful for the treatment of any cancer in which mis-match repair inhibition and / or cGAS / STING pathway activation is beneficial. Any suitable cancer may be targeted (e.g., adenoid cystic carcinoma, adrenal gland tumour, amyloidosis, anal cancer, appendix cancer, astrocytoma, ataxia-telangiectasia, Beckwith-Wiedemann Syndrome, bile duct cancer (cholangiocarcinoma), Birt-Hogg-Dube Syndrome, bladder cancer, bone cancer, brain stem glioma, brain tumour, oestrogen positive cancers, breast cancer, Carney Complex, central nervous system tumours, cervical cancer, colorectal cancer, Cowden Syndrome, craniopharyngioma, desmoplastic infantile ganglioglioma, ependymoma, esophageal cancer, Ewing sarcoma, eye cancer, eyelid cancer, familial adenomatous polyposis, familial GIST, familial malignant melanoma, familial non-VHL clear cell renal cell carcinoma, familial pancreatic cancer, gallbladder cancer, gastrointestinal stromal tumour- GIST, germ cell tumour, gestational trophoblastic disease, head and neck cancer, hereditary breast and ovarian cancer, hereditary diffuse gastric cancer, hereditary leiomyomatosis and renal cell cancer, hereditary mixed polyposis syndrome, hereditary pancreatitis, hereditary papillary renal carcinoma, juvenile polyposis syndrome, kidney cancer, lacrimal gland tumour, laryngeal and hypopharyngeal cancer, leukaemia (acute lymphoblastic leukaemia (ALL), acute myeloid leukaemia (AML), B-cell prolymphocytic leukaemia, hairy cell leukaemia, chronic lymphocytic leukaemia (CLL), chronic myeloid leukaemia (CML), chronic T-cell lymphocytic leukaemia, eosinophilic leukaemia), Li- Fraumeni Syndrome, liver cancer, lung cancer (non-small cell lung cancer, small cell lung cancer), Lymphoma (Hodgkin, non-Hodgkin), Lynch Syndrome, mastocytosis, medulloblastoma, melanoma, meningioma, mesothelioma, multiple endocrine neoplasia Type 1 & 2, multiple myeloma, MUTYH (or MYH)-associated polyposis, myelodysplastic syndromes (MDS), nasal cavity and paranasal sinus Cancer, nasopharyngeal Cancer, neuroblastoma, neuroendocrine tumours (e.g., of the gastrointestinal tract, lung or pancreas), neurofibromatosis Type 1 & 2, nevoid basal cell carcinoma syndrome, oral and oropharyngeal cancer, osteosarcoma, ovarian I fallopian tube I peritoneal cancer, pancreatic cancer, parathyroid cancer, penile cancer, Peutz-Jeghers Syndrome, pheochromocytoma, paraganglioma, pituitary gland tumour, pleuropulmonary blastoma, androgen receptor positive cancers, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma (e.g., Kaposi or soft tissue), skin cancer, small bowel cancer, stomach cancer, testicular cancer, thymoma and thymic carcinoma, thyroid cancer, tuberous sclerosis complex, uterine cancer, vaginal cancer, Von Hippel-Lindau syndrome, vulvar cancer, Waldenstrom’s macroglobulinemia, Werner syndrome, Wilms Tumour and xeroderma pigmentosum). Particular cancers of interest include haematological cancers such as lymphomas (including diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Burkitt lymphoma (BL) andangioimmunoblastic T-cell lymphoma (AITL)), leukaemias (including acute lymphoblastic leukaemia (ALL) and chronic myeloid leukaemia (CML)), multiple myeloma, breast cancer, non-small cell lung cancer (NSCLC), colorectal cancer, endometrial cancer, gastro-oesophageal cancer, neuroendocrine cancers, osteosarcomas, prostate cancer, pancreatic cancer, small intestine cancer, bladder cancer, rectal cancer, cholangiocarcinoma, CNS cancer, thyroid cancer, head and neck cancer, oesophageal cancer, and ovarian cancer.

[0041] Features, including optional, suitable, and preferred features in relation to one aspect of the invention may also be features, including optional, suitable and preferred features in relation to any other aspect of the invention.DETAILED DESCRIPTION OF THE INVENTIONDefinitions

[0042] Unless otherwise stated, the following terms used in the specification and claims have the following meanings set out below.

[0043] It is to be appreciated that references to “treating” or “treatment” include prophylaxis as well as the alleviation of established symptoms of a condition. “Treating” or “treatment” of a state, disorder or condition therefore includes: (1) preventing or delaying the appearance of clinical symptoms of the state, disorder or condition developing in a human that may be afflicted with or predisposed to the state, disorder or condition but does not yet experience or display clinical or subclinical symptoms of the state, disorder or condition; (2) inhibiting the state, disorder or condition (i.e., arresting, reducing or delaying the development of the disease or a relapse thereof (in case of maintenance treatment) or at least one clinical or subclinical symptom thereof); or (3) relieving or attenuating the disease (i.e., causing regression of the state, disorder or condition or at least one of its clinical or subclinical symptoms).

[0044] A “therapeutically effective amount” means the amount of a compound that, when administered to a mammal for treating a disease, is sufficient to effect such treatment for the disease. The "therapeutically effective amount" will vary depending on the compound, the disease and its severity and the age, weight, etc., of the mammal to be treated. It should be understood that in, for example, a human or other mammal, a therapeutically effective amount can be determined experimentally in a laboratory or clinical setting, or a therapeutically effective amount may be the amount required by the guidelines of the United States Food and Drug Administration (FDA) or equivalent foreign regulatory body, for the particular disease and subject being treated. It should be appreciated that determination of proper dosage forms, dosage amounts, and routes of administration is within the level of ordinary skill in the pharmaceutical and medical arts.

[0045] As used herein by themselves or in conjunction with another term or terms, “subject(s)” and “patient(s)”, refer to animals (e.g., mammals), particularly humans. Suitably, the “subject(s)” and “patient(s)” may be a non-human animal (e.g., livestock and domestic pets) or a human.

[0046] As used herein by itself or in conjunction with another term or terms, “pharmaceutically acceptable” refers to materials that are generally chemically and / or physically compatible with other ingredients (such as, for example, with reference to a formulation), and / or is generally physiologically compatible with the recipient (such as, for example, a subject) thereof.

[0047] The phrase “compound of the invention” means those compounds which are disclosed herein.

[0048] "About" when used herein in conjunction with a measurable value such as, for example, an amount or a period of time and the like, is meant to encompass reasonable variations of the value, for instance, to allow for experimental error in the measurement of said value.Compounds

[0049] According to a first aspect of the present invention there is provided a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound is selected from any one of the following:

[0050] Accordingly, the present invention provides a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound is selected from any one of the following: (E)- / V-(5-chloro-2-(5-chloro-2-hydroxy-4-methoxybenzoyl)isoindolin-4-yl)-4-(dimethylamino)but-2- enamide; / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)-6-(2-(dimethylamino)ethoxy)isoindolin-4-yl)- / V- methylacrylamide;(E)-4-(dimethylamino)- / V-(2-(6-hydroxy-4-methoxy-2,3-dimethylbenzoyl)isoindolin-4-yl)- / V-methylbut-2- enamide;(E)- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)- / V-methyl-4-(methylamino)but- 2-enamide;(E)- / V-(6-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)-5,6,7,8-tetrahydro-1 ,6-naphthyridin-3-yl)-4- (dimethylamino)- / V-methylbut-2-enamide;(E)-4-(dimethylamino)- / V-(2-(2-hydroxy-5-isopropyl-4-methoxybenzoyl)-3-methyl-1 ,2,3,4- tetrahydroisoquinolin-7-yl)- / V-methylbut-2-enamide;(E)- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-4-(diethylamino)but-2-enamide;(E)-4-amino- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)- / V-methylbut-2- enamide;(E)- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)- / V-methyl-4-(pyrrolidin-1-yl)but- 2-enamide;(E)- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-4-(pyrrolidin-1-yl)but-2- enamide;(E)- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)- / V-methyl-3-(1-methylpyrrolidin- 2-yl)acrylamide;(E)- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)- / V-methyl-3-(pyrrolidin-2- yl)acrylamide;(E)- / V-(5-chloro-2-(5-cyclopropyl-2-hydroxy-4-methoxybenzoyl)isoindolin-4-yl)-4-(dimethylamino)but-2- enamide;(E)- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-4-((2- fluoroethyl)(methyl)amino)but-2-enamide;(E)- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-4- (cyclopropyl(methyl)amino)but-2-enamide;(E)-A / -(5-chloro-2-(3-chloro-4-hydroxy-2-methylbenzoyl)isoindolin-4-yl)-4-(dimethylamino)-A / -methylbut-2- enamide;(E)- / V-(2-(3-Chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-3-(1-methylpyrrolidin-2- yl)acrylamide;(E)- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-3-(pyrrolidin-2-yl)acrylamide; (E)- / V-(2-(5-chloro-2-hydroxy-4-methoxybenzoyl)isoindolin-4-yl)-3-(pyrrolidin-2-yl)acrylamide;(E)- / V-(5-chloro-2-(5-chloro-2-hydroxy-4-methoxybenzoyl)isoindolin-4-yl)-3-(pyrrolidin-2-yl)acrylamide;(E)- / V-(5-chloro-2-(5-chloro-2-hydroxy-4-methoxybenzoyl)isoindolin-4-yl)-3-(1-methylpyrrolidin-2- yl)acrylamide;(E)- / V-(2-(3-chloro-2-fluoro-6-hydroxy-4-methoxybenzoyl)isoindolin-4-yl)-3-(1-methylpyrrolidin-2- yl)acrylamide;(E)- / V-(2-(3-chloro-6-hydroxy-2,4-dimethoxybenzoyl)isoindolin-4-yl)-3-(1-methylpyrrolidin-2-yl)acrylamide;(E)- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-4-(isopropylamino)but-2- enamide;(E)- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-4-(dimethylamino)- / V-ethylbut-2- enamide;(E)- / V-(2-(3-bromo-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-4-(dimethylamino)- / V- methylbut-2-enamide;(E)- / V-(2-(3-bromo-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-4-(dimethylamino)but-2- enamide;(E)- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-4-(ethyl(methyl)amino)- / V- methylbut-2-enamide;(E)- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-4-(ethyl(methyl)amino)but-2- enamide;(E)- / V-(2-(3-chloro-6-hydroxy-2,4-dimethoxybenzoyl)isoindolin-4-yl)-4-(dimethylamino)but-2-enamide; and (E)- / V-(5-chloro-2-(5-chloro-2-hydroxy-4-methoxybenzoyl)isoindolin-4-yl)-N-methyl-3-(1-methylpyrrolidin- 2-yl)acrylamide.

[0051] The present invention provides a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound, or pharmaceutically acceptable salt, hydrate or solvate thereof is selected from any one of Examples 1-21 described herein. The present invention also provides a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound, or pharmaceutically acceptable salt, hydrate or solvate thereof is selected from any one of Examples 1-22 described herein.

[0052] In an embodiment, the compound is selected from any one of the following:thereof.

[0053] In an embodiment, the compound is selected from any one of the following:pharmaceutically acceptable salt, hydrate or solvate thereof.

[0054] In an embodiment, the compound is selected from any one of the following:or a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0055] In an embodiment, the compound is selected from any one of the following:thereof.

[0056] In an embodiment, the compound is, or a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0057] In an embodiment, the compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0058] In an embodiment, the compoundor a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0059] In an embodiment, the compoundor a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0060] pharmaceutically acceptable salt, hydrate or solvate thereof. In some embodiments, the compound is apharmaceutically acceptable salt, hydrate or solvate thereof.

[0062] In an embodiment, the compound isor a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0063] In an embodiment, the compoundor a pharmaceutically acceptable salt, hydrate or solvate thereof. In some embodiments, the compound is a hydrochloric acid salt thereof

[0064] In an embodiment, the compoundpharmaceutically acceptable salt, hydrate or solvate thereof.

[0065] In an embodiment, the compoundpharmaceutically acceptable salt, hydrate or solvate thereof.

[0066] In an embodiment, the compound isor a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0067] In an embodiment, the compoundor a pharmaceutically acceptable salt, hydrate or solvate thereof. In some embodiments, the compound is a hydrochloric acid salt thereof

[0068] In an embodiment, the compound isor a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0069] In an embodiment, the compoundpharmaceutically acceptable salt, hydrate or solvate thereof.

[0070] In an embodiment, the compoundpharmaceutically acceptable salt, hydrate or solvate thereof.

[0071] In an embodiment, the compound isor a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0072] In an embodiment, the compoundor a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0073] In an embodiment, the compound isor a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0074] In an embodiment, the compoundor a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0075] In an embodiment, the compoundor pharmaceutically acceptable salt, hydrate or solvate thereof.

[0076] In an embodiment, the compoundor pharmaceutically acceptable salt, hydrate or solvate thereof.

[0077] In an embodiment, the compoundor a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0078] In an embodiment, the compoundor a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0079] In an embodiment, the compoundor a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0080] In an embodiment, the compound isor pharmaceutically acceptable salt, hydrate or solvate thereof.

[0081] In an embodiment, the compound isor a pharmaceutically acceptable salt, hydrate or solvate thereof. In some embodiments, the compound is a trifluoroacetic acid salt thereof

[0082] In an embodiment, the compound isor a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0083] In an embodiment, the compoundpharmaceutically acceptable salt, hydrate or solvate thereof. In some embodiments, the compound is a trifluoroacetic acid salt thereof

[0084] In an embodiment, the compoundpharmaceutically acceptable salt, hydrate or solvate thereof. In some embodiments, the compound is a trifluoroacetic acid salt thereof

[0085] In an embodiment, the compoundor a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0086] In an embodiment, the compoundor a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0087] A suitable pharmaceutically acceptable salt of a compound of the invention is, for example, an acid-addition salt of a compound of the invention which is sufficiently basic, for example, an acid-addition salt with, for example, an inorganic or organic acid, for example hydrochloric, hydrobromic, sulfuric, phosphoric, trifluoroacetic, formic, citric methane sulfonate or maleic acid. Suitably, the acid-addition salt is formed with hydrochloric acid or trifluoroacetic acid. Thus, the present invention suitably provides a compound as described herein, or a hydrochloric acid salt thereof, or a trifluoroacetic acid salt thereof. In addition, a suitable pharmaceutically acceptable salt of a compound of the invention which is sufficiently acidic is an alkali metal salt, for example a sodium or potassium salt, an alkaline earth metal salt, for example a calcium or magnesium salt, an ammonium salt or a salt with an organic base which affords a pharmaceutically acceptable cation, for example a salt with methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris-(2-hydroxyethyl)amine.

[0088] Compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are termed “isomers”. Isomers that differ in the arrangement of their atoms in space are termed “stereoisomers”. Stereoisomers that are not mirror images of one another are termed “diastereoisomers” and those that are non-superimposable mirror images of each other are termed “enantiomers”. When a compound has an asymmetric centre, for example, it is bonded to four different groups, a pair of enantiomers is possible. An enantiomer can be characterized by the absolute configuration of its asymmetric centre and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the manner in which the molecule rotates the plane of polarized light and designated as dextrorotatory or levorotatory (i.e., as (+) or (-)-isomers respectively). A chiral compound can exist as either individual enantiomer or as a mixture thereof. A mixture containing equal proportions of the enantiomers is called a “racemic mixture”.

[0089] The compounds of this invention may possess one or more asymmetric centres; such compounds can therefore be produced as individual (R)- or (S)-stereoisomers or as mixtures thereof. Unless indicated otherwise, the description or naming of a particular compound in the specification and claims is intended to include both individual enantiomers and mixtures, racemic or otherwise, thereof. The methods for the determination of stereochemistry and the separation of stereoisomers are well-known in the art (see discussion in Chapter 4 of “Advanced Organic Chemistry”, 4th edition J. March, John Wiley and Sons, New York, 2001), for example by synthesis from optically active starting materials or byresolution of a racemic form. Some of the compounds of the invention may have geometric isomeric centres (E- and Z- isomers).

[0090] It is to be understood that the present invention encompasses all optical, diastereoisomers and geometric isomers and mixtures thereof that possess activity.

[0091] The present invention also encompasses compounds of the invention as defined herein which comprise one or more isotopic substitutions. For example, H may be in any isotopic form, including1H,2H (D), and3H (T); C may be in any isotopic form, including12C,13C, and14C; and O may be in any isotopic form, including16O and18O; and the like.

[0092] It is also to be understood that certain compounds of the invention may exist in solvated as well as unsolvated forms such as, for example, hydrated forms. It is to be understood that the invention encompasses all such solvated forms that possess activity.

[0093] It is also to be understood that certain compounds of the invention may exhibit polymorphism, and that the invention encompasses all such forms that possess activity.

[0094] Compounds of the invention may exist in a number of different tautomeric forms and references to compounds of the invention include all such forms. For the avoidance of doubt, where a compound can exist in one of several tautomeric forms, and only one is specifically described or shown, all others are nevertheless embraced by the compounds of the invention. Examples of tautomeric forms include keto-, enol-, and enolate-forms, as in, for example, the following tautomeric pairs: keto / enol (illustrated below), imine / enamine, amide / imino alcohol, amidine / amidine, nitroso / oxime, thioketone / enethiol, and nitro / aci- nitro.keto enol enolate

[0095] Compounds of the invention containing an amine function may also form N-oxides. A reference herein to a compound of the invention that contains an amine function also includes the N-oxide. Where a compound contains several amine functions, one or more than one nitrogen atom may be oxidised to form an N-oxide. Particular examples of N-oxides are the N-oxides of a tertiary amine or a nitrogen atom of a nitrogen-containing heterocycle. N-Oxides can be formed by treatment of the corresponding amine with an oxidizing agent such as hydrogen peroxide or a per-acid (e.g., a peroxycarboxylic acid), see for example Advanced Organic Chemistry, by Jerry March, 4th Edition, Wiley Interscience. More particularly, N-oxides can be made by the procedure of L. W. Deady (Syn. Comm. 1977, 7, 509-514) in which the amine compound is reacted with m-chloroperoxybenzoic acid (mCPBA), for example, in an inert solvent such as dichloromethane.

[0096] The compounds of the invention may be administered in the form of a pro-drug which is broken down in the human or animal body to release a compound of the invention. A pro-drug may be used to alter the physical properties and / or the pharmacokinetic properties of a compound of the invention. A prodrug can be formed when the compound of the invention contains a suitable group or substituent to which a property-modifying group can be attached. Examples of pro-drugs include in vivo cleavable ester derivatives that may be formed at a carboxy group or a hydroxy group in a compound of the invention andin-vivo cleavable amide derivatives that may be formed at a carboxy group or an amino group in a compound of the invention.

[0097] Accordingly, the present invention includes those compounds of the invention, as defined hereinbefore, when made available by organic synthesis and when made available within the human or animal body by way of cleavage of a pro-drug thereof. Accordingly, the present invention includes those compounds of the invention that are produced by organic synthetic means and also such compounds that are produced in the human or animal body by way of metabolism of a precursor compound, that is a compound of the invention, may be a synthetically-produced compound or a metabolically-produced compound.

[0098] A suitable pharmaceutically acceptable pro-drug of a compound of the invention is one that is based on reasonable medical judgement as being suitable for administration to the human or animal body without undesirable pharmacological activities and without undue toxicity.

[0099] Various forms of pro-drug have been described, for example in the following documents:- a) Methods in Enzymology, Vol. 42, p. 309-396, edited by K. Widder, et al. (Academic Press, 1985); b) Design of Pro-drugs, edited by H. Bundgaard, (Elsevier, 1985); c) A Textbook of Drug Design and Development, edited by Krogsgaard-Larsen and H. Bundgaard, Chapter s “Design and Application of Pro-drugs”, by H. Bundgaard p. 113-191 (1991); d) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992); e) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77, 285 (1988); f) N. Kakeya, et al., Chem. Pharm. Bull., 32, 692 (1984); g) T. Higuchi and V. Stella, “Pro-Drugs as Novel Delivery Systems”, A.C.S. Symposium Series, Volume 14; and h) E. Roche (editor), “Bioreversible Carriers in Drug Design”, Pergamon Press, 1987.

[0100] A suitable pharmaceutically acceptable pro-drug of a compound of invention, that possesses a carboxy group is, for example, an in vivo cleavable ester thereof. An in vivo cleavable ester of a compound of the invention containing a carboxy group is, for example, a pharmaceutically acceptable ester which is cleaved in the human or animal body to produce the parent acid or parent alcohol. Suitable pharmaceutically acceptable esters for carboxy include (1 -6C)alkyl esters such as methyl, ethyl and tertbuty , (1-6C)alkoxymethyl esters such as methoxymethyl esters, (1-6C)alkanoyloxymethyl esters such as pivaloyloxymethyl esters, 3-phthalidyl esters, (3-8C)cycloalkylcarbonyloxy-(1-6C)alkyl esters such as cyclopentylcarbonyloxymethyl and 1 -cyclohexylcarbonyloxyethyl esters, 2-oxo-1 ,3-dioxolenylmethyl esters such as 5-methyl-2-oxo-1 ,3-dioxolen-4-ylmethyl esters and (1-6C)alkoxycarbonyloxy-(1-6C)alkyl esters such as methoxycarbonyloxymethyl and 1 -methoxycarbonyloxyethyl esters.

[0101] A suitable pharmaceutically acceptable pro-drug of a compound of the invention that possesses a hydroxy group is, for example, an in vivo cleavable ester or ether thereof. An in vivo cleavable ester or ether of a compound of the invention containing a hydroxy group is, for example, a pharmaceutically acceptable ester or ether which is cleaved in the human or animal body to produce the parent hydroxy compound. Suitable pharmaceutically acceptable ester forming groups for a hydroxy group include inorganic esters such as phosphate esters (including phosphoramidic cyclic esters). Further suitable pharmaceutically acceptable ester forming groups for a hydroxy group include (1-10C)alkanoyl groupssuch as acetyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl groups, (1- 10C)alkoxycarbonyl groups such as ethoxycarbonyl, N,N-(1-6C)2carbamoyl, 2-dialkylaminoacetyl and 2- carboxyacetyl groups. Examples of ring substituents on the phenylacetyl and benzoyl groups include aminomethyl, N-alkylaminomethyl, N,N-dialkylaminomethyl, morpholinomethyl, piperazin-1 -ylmethyl and 4-(1-4C)alkylpiperazin-1-ylmethyl. Suitable pharmaceutically acceptable ether forming groups for a hydroxy group include acyloxyalkyl groups such as acetoxymethyl and pivaloyloxymethyl groups.

[0102] A suitable pharmaceutically acceptable pro-drug of a compound of the invention that possesses a carboxy group is, for example, an in vivo cleavable amide thereof, for example an amide formed with an amine such as ammonia, a (1-4C)alkylamine such as methylamine, a [(1-4C)alkyl]2amine such as dimethylamine, / V-ethyl- / V-methylamine or diethylamine, a (1-4C)alkoxy-(2-4C)alkylamine such as 2-methoxyethylamine, a phenyl-(1-4C)alkylamine such as benzylamine and amino acids such as glycine or an ester thereof.

[0103] A suitable pharmaceutically acceptable pro-drug of a compound of the invention that possesses an amino group is, for example, an in vivo cleavable amide derivative thereof. Suitable pharmaceutically acceptable amides from an amino group include, for example an amide formed with (1-10C)alkanoyl groups such as an acetyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl groups. Examples of ring substituents on the phenylacetyl and benzoyl groups include aminomethyl, N- alkylaminomethyl, N,N-dialkylaminomethyl, morpholinomethyl, piperazin-1 -ylmethyl and 4-(1- 4C)alkyl)piperazin-1 -ylmethyl.

[0104] The in vivo effects of a compound of the invention may be exerted in part by one or more metabolites that are formed within the human or animal body after administration of a compound of the invention. As stated hereinbefore, the in vivo effects of a compound of the invention may also be exerted by way of metabolism of a precursor compound (a pro-drug).

[0105] Suitably, the present invention excludes any individual compounds not possessing the biological activity defined herein.Synthesis

[0106] The compounds of the present invention can be prepared by any suitable technique known in the art. Particular processes for the preparation of these compounds are described further in the accompanying examples.

[0107] In the description of the synthetic methods described herein and in any referenced synthetic methods that are used to prepare the starting materials, it is to be understood that all proposed reaction conditions, including choice of solvent, reaction atmosphere, reaction temperature, duration of the experiment and workup procedures, can be selected by a person skilled in the art.

[0108] It is understood by one skilled in the art of organic synthesis that the functionality present on various portions of the molecule must be compatible with the reagents and reaction conditions utilised.

[0109] It will be appreciated that during the synthesis of the compounds of the invention in the processes defined herein, or during the synthesis of certain starting materials, it may be desirable to protect certain substituent groups to prevent their undesired reaction. The skilled chemist will appreciate when such protection is required, and how such protecting groups may be put in place, and later removed.

[0110] For examples of protecting groups see one of the many general texts on the subject, for example, ‘Protective Groups in Organic Synthesis’ by Theodora Green (publisher: John Wiley & Sons). Protecting groups may be removed by any convenient method described in the literature or known to the skilled chemist as appropriate for the removal of the protecting group in question, such methods being chosen so as to effect removal of the protecting group with the minimum disturbance of groups elsewhere in the molecule.

[0111] Thus, if reactants include, for example, groups such as amino, carboxy or hydroxy it may be desirable to protect the group in some of the reactions mentioned herein.

[0112] By way of example, a suitable protecting group for an amino or alkylamino group is, for example, an acyl group, for example an alkanoyl group such as acetyl, an alkoxycarbonyl group, for example a methoxycarbonyl, ethoxycarbonyl or f-butoxycarbonyl group, an arylmethoxycarbonyl group, for example benzyloxycarbonyl, or an aroyl group, for example benzoyl. The deprotection conditions for the above protecting groups necessarily vary with the choice of protecting group. Thus, for example, an acyl group such as an alkanoyl or alkoxycarbonyl group or an aroyl group may be removed by, for example, hydrolysis with a suitable base such as an alkali metal hydroxide, for example lithium or sodium hydroxide. Alternatively, an acyl group such as a fe / Y-butoxycarbonyl group may be removed, for example, by treatment with a suitable acid as hydrochloric, sulfuric or phosphoric acid or trifluoroacetic acid and an arylmethoxycarbonyl group such as a benzyloxycarbonyl group may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon, or by treatment with a Lewis acid for example boron tris(trifluoroacetate). A suitable alternative protecting group for a primary amino group is, for example, a phthaloyl group which may be removed by treatment with an alkylamine, for example dimethylaminopropylamine, or with hydrazine.

[0113] A suitable protecting group for a hydroxy group is, for example, an acyl group, for example an alkanoyl group such as acetyl, an aroyl group, for example benzoyl, or an arylmethyl group, for example benzyl. The deprotection conditions for the above protecting groups will necessarily vary with the choice of protecting group. Thus, for example, an acyl group such as an alkanoyl or an aroyl group may be removed, for example, by hydrolysis with a suitable base such as an alkali metal hydroxide, for example lithium, sodium hydroxide or ammonia. Alternatively, an arylmethyl group such as a benzyl group may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon.

[0114] A suitable protecting group for a carboxy group is, for example, an esterifying group, for example a methyl or an ethyl group which may be removed, for example, by hydrolysis with a base such as sodium hydroxide, or for example a f-butyl group which may be removed, for example, by treatment with an acid, for example an organic acid such as trifluoroacetic acid, or for example a benzyl group which may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon.

[0115] Resins may also be used as a protecting group.

[0116] The methodology employed to synthesise the compounds of this invention may vary. Suitable processes for their preparation are described further in the accompanying Examples.

[0117] Once a compound of this invention has been synthesised by any one of the processes defined herein, the processes may then further comprise the additional steps of:(i) removing any protecting groups present;(ii) converting one compound of this invention into another compound of this invention;(iii) forming a pharmaceutically acceptable salt, hydrate or solvate thereof; and / or(iv) forming a prodrug thereof.

[0118] An example of (ii) above is when a compound of this invention is synthesised and then one or more of the various substituent groups be further reacted to change the nature of the group and provide an alternative compound of this invention.

[0119] The resultant compounds of this invention can be isolated and purified using techniques well known in the art.

[0120] The compounds of this invention may be synthesised by the synthetic routes shown in the Examples section below.Biological Activity

[0121] The biological assays described in the Examples section herein may be used to measure the pharmacological effects of the compounds of the present invention.

[0122] Although the pharmacological properties of the compounds of this invention vary with structural change, as expected, the compounds of the invention were found to be active in a PMS2 in vitro assay as described in the Examples section.Pharmaceutical Compositions

[0123] According to a further aspect of the present invention, there is provided a pharmaceutical composition comprising a compound of the invention as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, in admixture with a pharmaceutically acceptable diluent or carrier.

[0124] According to a further aspect of the present invention, there is provided a pharmaceutical composition comprising a compound of the invention as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable excipient or carrier.

[0125] The compositions of the invention may be in a form suitable for oral use (for example as tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), for topical use (for example as creams, ointments, gels, or aqueous or oily solutions or suspensions), for administration by inhalation (for example as a finely divided powder or a liquid aerosol), for administration by insufflation (for example as a finely divided powder) or for parenteral administration (for example as a sterile aqueous or oily solution for intravenous, subcutaneous, intramuscular, intraperitoneal or intramuscular dosing or as a suppository for rectal dosing).

[0126] The compositions of the invention may be obtained by conventional procedures using conventional pharmaceutical excipients, well known in the art. Thus, compositions intended for oral use may contain, for example, one or more colouring, sweetening, flavouring and / or preservative agents.

[0127] An effective amount I therapeutically effective amount of a compound of the present invention for use in therapy is an amount sufficient to treat or prevent a proliferative condition referred to herein, slow its progression and / or reduce the symptoms associated with the condition.

[0128] The amount of active ingredient that is combined with one or more excipients to produce a single dosage form will necessarily vary depending upon the individual treated and the particular route ofadministration. For example, a formulation intended for oral administration to humans will generally contain, for example, from 0.5 mg to 0.5 g of active agent (more suitably from 0.5 mg to 100 mg, for example from 1 mg to 30 mg) compounded with an appropriate and convenient amount of excipients which may vary from about 5 percent by weight to about 98 percent by weight of the total composition.

[0129] The size of the dose for therapeutic or prophylactic purposes of a compound of the invention will naturally vary according to the nature and severity of the conditions, the age and sex of the animal or patient, and the route of administration, according to well-known principles of medicine.

[0130] In using a compound of the invention for therapeutic or prophylactic purposes it will generally be administered so that a daily dose in the range, for example, 0.1 mg / kg to 75 mg / kg body weight is received, given if required in divided doses. In general, lower doses will be administered when a parenteral route is employed. Thus, for example, for intravenous or intraperitoneal administration, a dose in the range, for example, 0.1 mg / kg to 30 mg / kg body weight will generally be used. Similarly, for administration by inhalation, a dose in the range, for example, 0.05 mg / kg to 25 mg / kg body weight will be used. Oral administration may also be suitable, particularly in tablet form. Typically, unit dosage forms will contain about 0.5 mg to 0.5 g of a compound of this invention.Therapeutic Uses and Applications

[0131] The present invention provides compounds that function as inhibitors of PMS2 activity.

[0132] The compounds of the invention, or a pharmaceutically acceptable salt, hydrate or solvate thereof, therefore, have potential therapeutic uses in a variety of disease states in which the inhibition of PMS2 activity is beneficial.

[0133] The present invention therefore provides a method of treating a disease or disorder in which the inhibition PMS2 activity is beneficial in a patient in need of such treatment, said method comprising administering to said patient a therapeutically effective amount of a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein.

[0134] The present invention provides a method of inhibiting PMS2 activity, in vitro or in vivo, said method comprising contacting a cell with a therapeutically effective amount of a compound or a pharmaceutically acceptable salt, hydrate or solvate thereof as defined herein, or a pharmaceutical composition as defined herein.

[0135] The present invention provides a method of treating a disease or disorder in which PMS2 activity is implicated in a patient in need of such treatment, said method comprising administering to said patient a therapeutically effective amount of a compound or a pharmaceutically acceptable salt, hydrate or solvate thereof as defined herein, or a pharmaceutical composition as defined herein.

[0136] The present invention provides a method of treating a proliferative disorder in a patient in need of such treatment, said method comprising administering to said patient a therapeutically effective amount of a compound or a pharmaceutically acceptable salt, hydrate or solvate thereof as defined herein, or a pharmaceutical composition as defined herein.

[0137] The present invention provides a method of treating cancer in a patient in need of such treatment, said method comprising administering to said patient a therapeutically effective amount of a compound or a pharmaceutically acceptable salt, hydrate or solvate thereof as defined herein, or a pharmaceuticalcomposition as defined herein. In a particular embodiment, the cancer is human cancer.

[0138] The present invention provides a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein for use in therapy.

[0139] The present invention provides a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein for use as a medicament.

[0140] The present invention provides a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein, or a pharmaceutical composition as defined herein for use in the inhibition of PMS2 activity.

[0141] The present invention provides a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein, or a pharmaceutical composition as defined herein for use in the treatment of a disease or disorder in which PMS2 activity is implicated.

[0142] The present invention provides a compound or a pharmaceutically acceptable salt, hydrate or solvate thereof as defined herein, or a pharmaceutical composition as defined herein, for use in the treatment of a proliferative disorder.

[0143] The present invention provides a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein for use in the treatment of cancer. In a particular embodiment, the cancer is human cancer. In a particular embodiment, the cancer is human cancer, in particular oestrogen positive cancers, such as breast cancer, or androgen receptor positive cancers, such as prostate cancer.

[0144] The present invention provides a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein, or a pharmaceutical composition as defined herein for use in the treatment of a disease or disorder in which the inhibition of PMS2 activity is beneficial.

[0145] The present invention provides a use of a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein, or a pharmaceutical composition as defined herein in the manufacture of a medicament for the treatment of a proliferative disorder.

[0146] The present invention provides a use of a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein, or a pharmaceutical composition as defined herein in the manufacture of a medicament for the treatment of cancer.

[0147] The present invention provides a use of a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein, or a pharmaceutical composition as defined herein in the manufacture of a medicament for the inhibition of PMS2 activity.

[0148] The present invention provides a use of a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein, or a pharmaceutical composition as defined herein in the manufacture of a medicament for the treatment of a disease or disorder in which PMS2 activity is implicated.

[0149] The present invention provides a use of a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein, or a pharmaceutical composition as defined herein in the manufacture of a medicament for the treatment of a disease or disorder in which the inhibition of PMS2 activity is beneficial.

[0150] The term "proliferative disorder", “proliferative condition” and “proliferative disease” are usedinterchangeably herein and pertain to an unwanted or uncontrolled cellular proliferation of excessive or abnormal cells which is undesired, such as, neoplastic or hyperplastic growth, whether in vitro or in vivo.

[0151] In the above-outlined aspects of the invention, the proliferative disorder is suitably cancer, and the cancer is suitably a human cancer. In particular, the compounds of the present invention will be useful for the treatment of any cancer in which mis-match repair inhibition and / or cGAS / STING pathway activation is beneficial. Any suitable cancer may be targeted (e.g., adenoid cystic carcinoma, adrenal gland tumour, amyloidosis, anal cancer, appendix cancer, astrocytoma, ataxia-telangiectasia, Beckwith-Wiedemann Syndrome, bile duct cancer (cholangiocarcinoma), Birt-Hogg-Dube Syndrome, bladder cancer, bone cancer, brain stem glioma, brain tumour, oestrogen positive cancers, breast cancer, Carney Complex, central nervous system tumours, cervical cancer, colorectal cancer, Cowden Syndrome, craniopharyngioma, desmoplastic infantile ganglioglioma, ependymoma, esophageal cancer, Ewing sarcoma, eye cancer, eyelid cancer, familial adenomatous polyposis, familial GIST, familial malignant melanoma, familial non-VHL clear cell renal cell carcinoma, familial pancreatic cancer, gallbladder cancer, gastrointestinal stromal tumour- GIST, germ cell tumour, gestational trophoblastic disease, head and neck cancer, hereditary breast and ovarian cancer, hereditary diffuse gastric cancer, hereditary leiomyomatosis and renal cell cancer, hereditary mixed polyposis syndrome, hereditary pancreatitis, hereditary papillary renal carcinoma, juvenile polyposis syndrome, kidney cancer, lacrimal gland tumour, laryngeal and hypopharyngeal cancer, leukemia (acute lymphoblastic leukamia (ALL), acute myeloid leukemia (AML), B- cell prolymphocytic leukemia, hairy cell leukemia, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic T-cell lymphocytic leukemia, eosinophilic leukemia), Li-Fraumeni Syndrome, liver cancer, lung cancer (non-small cell lung cancer, small cell lung cancer), Lymphoma (Hodgkin, nonHodgkin), Lynch Syndrome, mastocytosis, medulloblastoma, melanoma, meningioma, mesothelioma, multiple endocrine neoplasia Type 1 & 2, multiple myeloma, MUTYH (or MYH)-associated polyposis, myelodysplastic syndromes (MDS), nasal cavity and paranasal sinus Cancer, nasopharyngeal Cancer, neuroblastoma, neuroendocrine tumours (e.g., of the gastrointestinal tract, lung or pancreas), neurofibromatosis Type 1 & 2, nevoid basal cell carcinoma syndrome, oral and oropharyngeal cancer, osteosarcoma, ovarian I fallopian tube I peritoneal cancer, pancreatic cancer, parathyroid cancer, penile cancer, Peutz-Jeghers Syndrome, pheochromocytoma, paraganglioma, pituitary gland tumour, pleuropulmonary blastoma, androgen receptor positive cancers, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma (e.g., Kaposi or soft tissue), skin cancer, small bowel cancer, stomach cancer, testicular cancer, thymoma and thymic carcinoma, thyroid cancer, tuberous sclerosis complex, uterine cancer, vaginal cancer, Von Hippel-Lindau syndrome, vulvar cancer, Waldenstrom’s macroglobulinemia, Werner syndrome, Wilms Tumour and xeroderma pigmentosum). Particular cancers of interest include haematological cancers such as lymphomas (including diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), Burkitt lymphoma (BL) and angioimmunoblastic T- cell lymphoma (AITL)), leukaemias (including acute lymphoblastic leukaemia (ALL) and chronic myeloid leukaemia (CML)), multiple myeloma, breast cancer, non-small cell lung cancer (NSCLC), colorectal cancer, endometrial cancer, gastro-oesophageal cancer, neuroendocrine cancers, osteosarcomas, prostate cancer, pancreatic cancer, small intestine cancer, bladder cancer, rectal cancer, cholangiocarcinoma, CNS cancer, thyroid cancer, head and neck cancer, oesophageal cancer, andovarian cancer.

[0152] The compounds of the present invention may also be used to treat triplet repeat disorders.

[0153] Thus, a further aspect of the present invention provides a method of treating a triplet repeat disorder in a patient in need of such treatment, said method comprising administering to said patient a therapeutically effective amount of a compound or a pharmaceutically acceptable salt, hydrate or solvate thereof as defined herein, or a pharmaceutical composition as defined herein. In a particular embodiment, the triplet repeat disorder is selected from the group consisting of Huntington’s disease (HD), myotonic dystrophy type 1 (DM1), fragile X syndrome type A (FRAXA), Friedreich’s ataxia (FRDA), and spinocerebellar ataxias (SCAs).

[0154] According to a further aspect of the present invention, there is provided a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein for use in the treatment of a triplet repeat disorder. In a particular embodiment, the triplet repeat disorder is selected from the group consisting of Huntington’s disease (HD), myotonic dystrophy type 1 (DM1), fragile X syndrome type A (FRAXA), Friedreich’s ataxia (FRDA), and spinocerebellar ataxias (SCAs).

[0155] According to a further aspect of the present invention, there is provided the use of a compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein, or a pharmaceutical composition as defined herein in the manufacture of a medicament for the treatment of a triplet repeat disorder. In a particular embodiment, the triplet repeat disorder is selected from the group consisting of Huntington’s disease (HD), myotonic dystrophy type 1 (DM1), fragile X syndrome type A (FRAXA), Friedreich’s ataxia (FRDA), and spinocerebellar ataxias (SCAs).Routes of Administration

[0156] The compounds of the invention or pharmaceutical compositions comprising these compounds may be administered to a subject by any convenient route of administration, whether systemically, peripherally or topically (i.e., at the site of desired action).

[0157] Routes of administration include, but are not limited to, oral (e.g., by ingestion); buccal; sublingual; transdermal (e.g., by a patch, plaster, etc.); transmucosal (e.g., by a patch, plaster, etc.); intranasal (e.g., by nasal spray); ocular (e.g., by eye drops); pulmonary (e.g., by inhalation or insufflation therapy using, e.g., via an aerosol, e.g., through the mouth or nose); rectal (e.g., by suppository or enema); vaginal (e.g., by pessary); parenteral, (e.g., by injection, including intratumoural, subcutaneous, intradermal, intramuscular, intravenous, intra-arterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal, subcuticular, intraarticular, subarachnoid, and intrasternal); by implant of a depot or reservoir (e.g., subcutaneously or intramuscularly).Combination Therapies

[0158] The compounds of the present invention may be administered as a sole therapy or may involve, in addition to a compound of the invention, conventional surgery or radiotherapy or chemotherapy or a targeted agent. Such chemotherapy or targeted agent may include one or more of the following categories:(i) Antiproliferative / antineoplastic drugs and combinations thereof, as used in medical oncology, such as, but not limited to, alkylating agents (for example cis-platin, oxaliplatin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulphan, temozolamide and nitrosoureas); antimetabolites (for example gemcitabine and antifolates such as fluoropyrimidines like 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside, and hydroxyurea); antitumour antibiotics (for example anthracyclines like adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin and mithramycin); antimitotic agents (for example vinca alkaloids like vincristine, vinblastine, vindesine and vinorelbine and taxoids like taxol and taxotere and polokinase inhibitors); and topoisomerase inhibitors (for example epipodophyllotoxins like etoposide and teniposide, amsacrine, topotecan and camptothecins including irinotecan);(ii) cytostatic agents such as, but not limited to, antioestrogens (for example tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene and iodoxyfene), antiandrogens (for example bicalutamide, flutamide, nilutamide and cyproterone acetate), LHRH antagonists or LHRH agonists (for example goserelin, leuprorelin and buserelin), steroid hormones, including progestogens (for example megestrol acetate) and corticosteroids (for example dexamethasone, prednisone and prednisolone), aromatase inhibitors (for example as anastrozole, letrozole, vorazole and exemestane) and inhibitors of 5a-reductase such as finasteride;(iii) anti-invasion agents such as, but not limited to, c-Src kinase family inhibitors 4-(6-chloro-2,3- methylenedioxyanilino)-7-[2-(4-methylpiperazin-1-yl)ethoxy]-5-tetrahydropyran-4-yloxyquinazoline (AZD0530; International Patent Application WO 01 / 94341), / V-(2-chloro-6-methylphenyl)-2-{6-[4-(2- hydroxyethyl)piperazin-1-yl]-2-methylpyrimidin-4-ylamino}thiazole-5-carboxamide (dasatinib, BMS- 354825; J. Med. Chem., 2004, 47, 6658-6661), bosutinib (SKI-606), and metalloproteinase inhibitors such as marimastat, inhibitors of urokinase plasminogen activator receptor function or antibodies to Heparanase;(iv) inhibitors of growth factor function such as, but not limited to, growth factor antibodies and growth factor receptor antibodies (for example the anti-erbB2 antibody trastuzumab [Herceptin™], the anti- EGFR antibody panitumumab, the anti-erbB1 antibody cetuximab [Erbitux, C225] and any growth factor or growth factor receptor antibodies disclosed by Stern et al. (Critical reviews in oncology / haematology, 2005, Vol. 54, pp11-29); such inhibitors also include tyrosine kinase inhibitors, for example inhibitors of the epidermal growth factor family (for example EGFR family tyrosine kinase inhibitors such as / V-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3- morpholinopropoxy)quinazolin-4-amine (gefitinib, ZD1839), / V-(3-ethynylphenyl)-6,7-bis(2- methoxyethoxy)quinazolin-4-amine (erlotinib, OSI-774) and 6-acrylamido- / V-(3-chloro-4- fluorophenyl)-7-(3-morpholinopropoxy)-quinazolin-4-amine (Cl 1033), erbB2 tyrosine kinase inhibitors such as lapatinib); inhibitors of the hepatocyte growth factor family; inhibitors of the insulin growth factor family; inhibitors of the platelet-derived growth factor family such as imatinib and / or nilotinib (AMN107); inhibitors of serine / threonine kinases (for example Ras / Raf signalling inhibitors such as farnesyl transferase inhibitors, for example sorafenib (BAY 43-9006), tipifarnib (R1 15777) and lonafarnib (SCH66336)), inhibitors of cell signalling through MEK and / or AKT kinases, c-kitinhibitors, abl kinase inhibitors, PI3 kinase inhibitors, Plt3 kinase inhibitors, CSF-1 R kinase inhibitors, IGF receptor (insulin-like growth factor) kinase inhibitors; aurora kinase inhibitors and cyclin dependent kinase inhibitors such as CDK2 and / or CDK4 inhibitors;(v) antiangiogenic agents such as, but not limited to, those which inhibit the effects of vascular endothelial growth factor, for example the anti-vascular endothelial cell growth factor antibody bevacizumab (Avastin™) and for example, a VEGF receptor tyrosine kinase inhibitor such as vandetanib (ZD6474), vatalanib (PTK787), sunitinib (SU11248), axitinib (AG-013736) and pazopanib (GW 786034);(vi) vascular damaging agents such as, but not limited to, Combretastatin A4 and compounds disclosed in International Patent Applications WO 99 / 02166, WO 00 / 40529, WO 00 / 41669, WO 01 / 92224, WO 02 / 04434 and WO 02 / 08213;(vii) an endothelin receptor antagonist, for example zibotentan (ZD4054) or atrasentan;(viii) antisense therapies, such as, but not limited to, those directed to targets listed above, such as ISIS 2503, an anti-ras antisense;(ix) immunotherapy approaches, including for example cancer vaccines, antibody, viral (oncolytic viruses) and small molecule or cell therapy approaches to increase the immunogenicity of patient tumour cells and / or facilitate a cell mediated anti-tumour response. Such therapies could include, but are not limited to, 0X40 agonists, cGAS-STING agonists, ENPP1 inhibitors, CD38 inhibitors, TBK1 inhibitors, A2a receptor antagonists, PI3 kinase inhibitors, TLR7 / 8 agonists, IDO inhibitors, Arginase inhibitors, BTK inhibitors and Bromodomain inhibitors; transduction with microbial vectors of cancer antigens, direct transduction of cancer antigens into antigen presenting cells, treatment with immune cells specific for cancer antigens (e.g., CAR-T), treatment with antibodies, antibody fragments and antibody drug conjugates that enable the immune system to recognise tumour cells.

[0159] Such conjoint treatment may be achieved by way of the simultaneous, sequential or separate dosing of the individual components of the treatment. Such combination products employ the compounds of the invention within the dosage ranges defined herein and the other pharmaceutically-active agent within its approved dosage range.

[0160] In one aspect, the present invention provides a combination for use in the treatment of a proliferative disorder, such as cancer (for example a cancer involving a solid tumour), comprising a compound of the invention as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, and any one of the anti-tumour agents defined herein. In an embodiment, the proliferative disorder is cancer.

[0161] In one aspect, the present invention provides a method of treating a proliferative disorder, such as cancer (for example a cancer involving a solid tumour), said method comprising administering to a subject a compound of the invention as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, and any one of the anti-tumour agents defined herein. In an embodiment, the proliferative disorder is cancer.

[0162] In one aspect, the present invention provides a use of a combination comprising a compound of the invention as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, and any one of the anti-tumour agents defined herein in the manufacture of a medicament for the treatment of aproliferative disorder, such as cancer (for example a cancer involving a solid tumour). In an embodiment, the proliferative disorder is cancer.

[0163] In a further aspect of the invention there is provided a compound of the invention or a pharmaceutically acceptable salt, hydrate or solvate thereof, for use in the treatment of cancer in combination with another anti-tumour agent, optionally selected from one defined herein.

[0164] In a further aspect of the invention there is provided a method of treating cancer, said method comprising administering to a subject a compound of the invention or a pharmaceutically acceptable salt, hydrate or solvate thereof in combination with another anti-tumour agent, optionally selected from one defined herein.

[0165] In a further aspect of the invention there is provided a use of a compound of the invention or a pharmaceutically acceptable salt, hydrate or solvate thereof in combination with another anti-tumour agent, optionally selected from one defined herein, in the manufacture of a medicament for the treatment of cancer.

[0166] In a further aspect of the invention there is provided a compound of the invention or a pharmaceutically acceptable salt, hydrate or solvate thereof, for use in the treatment of cancer in combination with a tyrosine kinase inhibitor, optionally selected from one defined herein.

[0167] In a further aspect of the invention there is provided method of treating cancer, said method comprising administering to a subject a compound of the invention or a pharmaceutically acceptable salt, hydrate or solvate thereof in combination with a tyrosine kinase inhibitor, optionally selected from one defined herein.

[0168] In a further aspect of the invention there is provided a use of a compound of the invention or a pharmaceutically acceptable salt, hydrate or solvate thereof in combination with a tyrosine kinase inhibitor, optionally selected from one defined herein, in the manufacture of a medicament for the treatment of cancer.

[0169] Herein, where the term “combination” is used it is to be understood that this refers to simultaneous, separate or sequential administration. In one aspect of the invention “combination” refers to simultaneous administration. In another aspect of the invention “combination” refers to separate administration. In a further aspect of the invention “combination” refers to sequential administration. Where the administration is sequential or separate, the delay in administering the second component should not be such as to lose the beneficial effect of the combination.

[0170] According to a further aspect of the invention there is provided a pharmaceutical composition which comprises a compound of the invention, or a pharmaceutically acceptable salt, hydrate or solvate thereof, in combination with an anti-tumour agent (optionally selected from one defined herein), in admixture with a pharmaceutically acceptable diluent or carrier.

[0171] According to a further aspect of the invention there is provided a pharmaceutical composition which comprises a compound of the invention, or a pharmaceutically acceptable salt, hydrate or solvate thereof, in combination with an anti-tumour agent (optionally selected from one defined herein), and a pharmaceutically acceptable excipient or carrier.Combination Therapy with Immune Modulating TreatmentsImmune checkpoint inhibitors

[0172] Immune checkpoint proteins present on immune cells and / or cancer cells [e.g., CTLA4 (also known as cytotoxic T-lymphocyte-associated protein 4 and CD152), LAG3 (also known as lymphocyteactivation gene 3 and CD223), PD1 (also known as programmed cell death protein 1 and CD279), PD-L1 (also known as programmed death-ligand 1 and CD274), TIM-3 (also known as T-cell immunoglobulin mucin-3) and TIGIT (also known as T-cell Immunoreceptor with Ig and ITIM domains)] are molecular targets that have been found to play an important role in regulating anti-tumour immune responses. Inhibitors of these immune checkpoint proteins (e.g., CTLA4, LAG3, PD1 , PD-L1 , TIM-3 and / or TIGIT inhibitors) promote an anti-tumour immune response that can be utilised to effectively treat certain forms of cancer.Immune stimulators

[0173] Monoclonal antibodies, bispecific antibodies, recombinant ligands and small molecule therapeutics that bind to stimulatory receptors on immune cells can facilitate an effective anti-tumour response. Such receptors may be involved in cell-to-cell contact for example contact between tumour cell and immune cell or between two types of immunce cells, other receptors may bind to soluble factors that stimulate an immune response. In one such embodiment antibodies, bispecifics, recombindant proteins or small molecule therapeutics can activate stimulatory receptors, including, but not limited to, 4-1 BB, 0X40, cGAS-STING, CD27, CD40, and DR3 that enhance anti-tumour immunity.

[0174] Modulators of antigen processing may facilitate the presentation of neoantigenic peptides on the cell surface to enhance an effective anti-tumour response. In one such embodiment inhibitors of the endoplasmic reticulum aminopeptidases ERAP1 and ERAP2 may stimulate anti-tumour immunity.

[0175] In one aspect, the present invention relates to a combination comprising a compound as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, and an immune checkpoint inhibitor or immune stimulator as defined herein, or a pharmaceutically acceptable salt thereof, for use in the treatment of a proliferative disorder.

[0176] In another aspect, the present invention relates to a use of a combination comprising a compound as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, and an immune checkpoint inhibitor or immune stimulator as defined herein, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treatment of a proliferative disorder.

[0177] In another aspect, the present invention relates to a method of treating a proliferative disorder in a subject in need thereof comprising administering to said subject a combination comprising a compound as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, and an immune checkpoint inhibitor or immune stimulator as defined herein, or a pharmaceutically acceptable salt thereof, as defined herein.

[0178] In another aspect, the present invention relates to a compound as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein for use in the treatment of a proliferative disorder, wherein the compound, or a pharmaceutically acceptable salt thereof, is for simultaneous, separate or sequential administeration with an immune checkpoint inhibitor, or immune stimulator, or a pharmaceutically acceptable salt thereof.

[0179] In another aspect, the present invention relates to an immune checkpoint inhibitor or immune stimulator, or a pharmaceutically acceptable salt thereof, for use in the treatment of a proliferative disorder, wherein the immune checkpoint inhibitor is for simultaneous, separate or sequential administeration with a compound as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein.

[0180] In another aspect, the present invention relates to a use of a compound as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein in the manufacture of a medicament for treating a proliferative disorder, wherein the medicament is for simultaneous, separate or sequential administeration with an immune checkpoint inhibitor or immune stimulator, or a pharmaceutically acceptable salt thereof.

[0181] In another aspect, the present invention relates to a use of an immune checkpoint inhibitor or immune stimulator, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating a proliferative disorder, wherein the medicament is for simultaneous, separate or sequential administeration with a compound as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0182] In another aspect, the present invention relates to a method of treating a proliferative disorder comprising adminstering to a subject in need thereof a therapeutically effective amount of a compound as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein and an immune checkpoint inhibitor or immune stimulator as defined herein, or a pharmaceutically acceptable salt thereof, either sequentially, separately or simultaneously.

[0183] Any immune checkpoint inhibitor or immune stimulator may be used in the combination therapy defined herein.

[0184] In one embodiment, the immune stimulator is selected from a 4-1 BB stimulator, a 0X40 stimulator, a CD27 stimulator, a CD40 stimulator, and a DR3 stimulator. In another embodiment the immune checkpoint inhibitor is selected from a PD1 -inhibitor, a PD-L1 inhibitor, a LAG3 inhibitor, CTLA-4 inhibitor, a TIM-3 inhibitor and / or a TIGIT inhibitor. In a particular embodiment, the immune checkpoint inhibitor is a PD1 or PD-L1 inhibitor.

[0185] PD-1 is a cell surface receptor protein present on immune cells such as T cells. PD-1 plays an important role in down-regulating the immune system and promoting self-tolerance by suppressing T cell activation. The PD-1 protein is an immune checkpoint that guards against autoimmunity through a dual mechanism of promoting apoptosis (programmed cell death) in antigen specific T cells in lymph nodes, while simultaneously reducing apoptosis in regulatory T cells (anti-inflammatory suppressive T cells).

[0186] PD-1 therefore inhibits the immune system. This prevents autoimmune diseases, but it can also prevent the immune system from killing cancer cells.

[0187] PD1 binds two ligands, PD-L1 and PD-L2. PD-L1 is of particular interest as it is highly expressed in several cancers and hence the role of PD1 in cancer immune evasion is well established. Monoclonal antibodies targeting PD-1 that boost the immune system are approved or are being developed for the treatment of cancer. Many tumour cells express PD-L1 , an immunosuppressive PD-1 ligand; inhibition of the interaction between PD-1 and PD-L1 can enhance T-cell responses in vitro and mediate preclinical antitumour activity. This is known as immune checkpoint blockade.

[0188] Examples of drugs that target PD-1 include pembrolizumab (Keytruda) and nivolumab (Opdivo). These drugs have been shown to be effective in treating several types of cancer, including melanoma of the skin, non-small cell lung cancer, kidney cancer, bladder cancer, head and neck cancers, and Hodgkin lymphoma. They are also being studied for use against many other types of cancer. Examples of drugs in development include BMS-936559 (Bristol Myers Squibb), MGA012 (MacroGenics) and MEDI-0680 (Medlmmune).

[0189] Examples of drugs that inhibit PD-L1 include atezolizumab (Tecentriq), avelumab (Bavencio) and durvalumab (Imfinzi). These drugs have also been shown to be helpful in treating different types of cancer, including bladder cancer, non-small cell lung cancer, and Merkel cell skin cancer (Merkel cell carcinoma). They are also being studied for use against other types of cancer.

[0190] Examples of LAG 3 inhibitors include BMS-986016 / Relatlimab, TSR-033, REGN3767, MGD013 (bispecific DART binding PD-1 and LAG-3), GSK2831781 and LAG525.

[0191] Examples of CTLA-4 inhibitors include MDX-010 / lpilimumab, AGEN1884, and CP- 675, 206 / Tremelimumab.

[0192] Examples of TIM-3 inhibitors include MBG453 (Novartis), TSR-022 (Tesaro), and LY3321367 (Lilly).

[0193] Examples of TIGIT inhibitors include Tiragolumab (MTIG7192A; RG6058; Genentech / Roche), AB154 (Arcus Bioscience), MK-7684 (Merck), BMS-986207 (Bristol-Myers Squibb), ASP8374 (Astellas Pharma; Potenza Therapeutics).

[0194] In one embodiment, the immune checkpoint inhibitor is selected from BMS-986016 / Relatlimab, TSR-033, REGN3767, MGD013 (bispecific DART binding PD-1 and LAG-3), GSK2831781 , LAG525, MDX-010 / lpilimumab, AGEN1884, and CP-675, 206 / Tremelimumab, pembrolizumab, nivolumab, atezolizumab, avelumab, durvalumab, MBG453, TSR-022, LY3321367, Tiragolumab (MTIG7192A; RG6058), AB154, MK-7684, BMS-986207, and / or ASP8374 or a pharmaceutically acceptable salt or solvate thereof.Combination therapy with DNA damage response modulators

[0195] The compounds of the present invention are particularly suited to use in combination with agents that act as DNA damage response modulators, e.g., PARP inhibitors, ATM inhibitors and ATR inhibitors.

[0196] In one aspect, the present invention relates to a combination comprising a compound as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a DNA damage response modulator (e.g., a PARP inhibitor, an ATM inhibitor and / or an ATR inhibitor), or a pharmaceutically acceptable salt thereof, for use in the treatment of a proliferative disorder.

[0197] In another aspect, the present invention relates to a use of a combination comprising a compound as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a DNA damage response modulator (e.g., a PARP inhibitor, an ATM inhibitor and / or an ATR inhibitor), or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating of a proliferative disorder.

[0198] In another aspect, the present invention relates to a method of treating of a proliferative disorder in a subject in need thereof comprising administering to said subject a combination comprising a compoundas defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a DNA damage response modulator (e.g., a PARP inhibitor, an ATM inhibitor and / or an ATR inhibitor), or a pharmaceutically acceptable salt thereof, as defined herein.

[0199] In another aspect, the present invention relates to a compound as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein for use in the treatment of a proliferative disorder, wherein the compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, is for simultaneous, separate or sequential administeration with a DNA damage response modulator (e.g., a PARP inhibitor, an ATM inhibitor and / or an ATR inhibitor), or a pharmaceutically acceptable salt thereof.

[0200] In another aspect, the present invention relates to a use of a compound as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein in the manufacture of a medicament for treating a proliferative disorder, wherein the medicament is for simultaneous, separate or sequential administeration with a DNA damage response modulator (e.g., a PARP inhibitor, an ATM inhibitor and / or an ATR inhibitor), or a pharmaceutically acceptable salt thereof.

[0201] In another aspect, the present invention relates to a method of treating a proliferative disorder comprising adminstering to a subject in need thereof a therapetuically effective amount of a compound as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, as defined herein and a DNA damage response modulator (e.g., a PARP inhibitor, an ATM inhibitor and / or an ATR inhibitor), or a pharmaceutically acceptable salt thereof, either sequentially, separately or simultaneously.

[0202] Any DNA damage response modulator (e.g., a PARP inhibitor, an ATM inhibitor and / or an ATR inhibitor) may be used in the combination therapy defined herein.EXAMPLES

[0203] While specific embodiments of the invention have been described herein for the purpose of reference and illustration, various modifications will be apparent to a person skilled in the art without departing from the scope of the invention as defined by the appended claims.AbbreviationsACN AcetonitrileCPME Cyclopentyl methyl etherDCM DichloromethaneDIPEA DIPEADMF / V, / V-DimethylformamideDMSO Dimethyl sulfoxideEDC / -(3-Dimethylaminopropyl)- / '-ethylcarbodiimide hydrochlorideEtOAc Ethyl acetateEtOH EthanolHATU Hexafluorophosphate Azabenzotriazole Tetramethyl UraniumHCI Hydrochloric acidHOBt 1 -Hydroxybenzotriazole hydrateMeOH MethanolNCS / V-ChlorosuccinimideNBS N-BromosuccinimideNMM N-MethylmorpholineRT Retention time or room temperatureSFC Supercritical Fluid ChromatographySTAB Sodium triacetoxyborohydrideTEA TriethylamineTFA Trifluoroacetic acidTHF TetrahydrofuranAnalytical Methods

[0204] Commercially available starting materials, reagents and dry solvents were used as supplied.Liquid Chromatography-Mass Spectrometry MethodsLC-Method 1

[0205] UPLC-MS was performed on a Waters DAD + Waters SQD2, single quadrupole UPLC-MS spectrometer using an Acquity UPLC BEH C18 1 ,7um 100 x 2.1 mm (Plus guard cartridge), maintained at 40°C column being initially held at 5% Acetonitrile (Far UV grade) with 0.1 % (V / V) formic acid / Water (High purity via PureLab Option unit) with 0.1 % formic acid for 0.4 minutes, followed by a linear gradient of 5- 95% within 6.4 minutes and then held at 95% for 1 .2 minutes (F = 0.4 mL / min).Method A

[0206] Waters Acquity UPLC with binary solvent manager, PDA detector and Acquity QDA performance mass detector, column: X-Bridge BEH C18, 50 x 2.1 mm, 2.5 micron, column temperature: 35°C, auto sampler temperature: 5°C, mobile phase A : 0.1 % (v / v) formic acid in water (pH = 2.70), Mobile Phase B : 0.1 % formic acid (v / v) in water : acetonitrile (10:90), mobile phase gradient details: t = 0 min (97% A, 3% B) flow : 0.8 mL / min; t = 0.75 min (97% A, 3% B) flow : 0.8 mL / min; gradient to t = 2.7 min (2% A, 98% B) flow : 0.8 mL / min; gradient to t = 3 min (0% A, 100% B) flow : 1 mL / min; t = 3.5 min (0% A, 100% B) flow : 1 mL / min; gradient to t = 3.51 min (97% A, 3% B) flow : 0.8 mL / min; end of run at t = 4 min (97% A, 3% B), Flow rate: 0.8 mL / min, analysis time 4 min. Mass detector parameter: ionization mode was cycled through positive and negative modes with cone voltage 10 V and 30 V and 0.8 kV capillary voltage, temperature of source and probe were 120°C and 600°C respectively.Method N

[0207] Waters Acquity UPLC- H Class equipped with PDA and attached with QDa detector, column: X- Bridge BEH C18, 50 x 2.1 mm, 2.5 micron, Column temperature: Ambient, Auto sampler temperature: 15°C, Mobile Phase A : 2 mM ammonium acetate followed by 0.1 %Formic acid in water, Mobile Phase B : 0.1 % Formic Acid in Acetonitrile, Mobile phase gradient details: T = 0 min (95% A, 5% B) flow; T = 0.4 min (95% A, 5% B) ; gradient to T = 0.8 min (65% A, 35% B) ; gradient to T = 1 .20 min (45% A, 55% B) ; T = 2.5 min (0% A, 100% B) ; gradient to T= 3.30 min (0% A, 100% B) ; gradient to T= 3.31 min to end of run at T = 4 min (95% A, 5% B), Flow rate: 0.55 mL / min, Run Time: 4 min. UV Detection Method:PDA Mass parameter: Probe:ESI, Mode of Ionisation : positive and negative, Cone voltage : 10V and 30V, capillary voltage: 0.8 KV, Extractor Voltage: 1 KV, Rf Lens: 0.1 .Temperature of source: 120°C, Temperature of Probe: 600°C, Cone Gas Flow:- Default, Desolvation Gas flow:-Default.Method O

[0208] Waters Acquity UPLC- H Class equipped with PDA and attached with QDa detector, column: X- Bridge BEH C18, 50 x 2.1 mm, 2.5 micron, Column temperature: Ambient, Auto sampler temperature: 15°C, Mobile Phase A : 5mM Ammonium Bicarbonate in Milli Q water, Mobile Phase B : Acetonitrile, Mobile phase gradient details: T = 0 min (95% A, 5% B) flow; T = 0.4 min (95% A, 5% B) ; gradient to T = 0.8 min (65% A, 35% B) ; gradient to T = 1 .20 min (45% A, 55% B) ; T = 2.5 min (0% A, 100% B) ; gradient to T= 3.30 min (0% A, 100% B) ; gradient to T= 3.31 min to end of run at T = 4 min (95% A, 5% B), Flow rate: 0.55 mL / min, Run Time:- 4 min. UV Detection Method: PDA Mass parameter: Probe:ESI, Mode of Ionisation positive and negative, Cone voltage : 10V and 30V, capillary voltage: 0.8 KV, Extractor Voltage: 1 KV, Rf Lens: 0.1 .Temperature of source: 120°C, Temperature of Probe: 600°C, Cone Gas Flow:- Default, Desolvation Gas flow:-Default.NMR

[0209] 1H Nuclear magnetic resonance (NMR) spectroscopy was carried out using a Bruker instrument operating at 400 MHz using the stated solvent at around room temperature unless otherwise stated. In all cases, NMR data were consistent with the proposed structures. Characteristic chemical shifts (6) are given in parts-per-million using conventional abbreviations for designation of major peaks: e.g. s, singlet; d, doublet; t, triplet; q, quartet; dd, doublet of doublets; dt, doublet of triplets; m, multiplet; br, broad.Analytical HPLC MethodsMethod: H

[0210] Machine Details: - Shimadzu LC-2050-C with PDA detector (HP-09), Waters alliance e2695 with 2998 PDA detector (HP-11) and Agilent Infinity II G6125C (HP-05), Column temperature: 25°C, Auto sampler temperature: 25°C, Mobile Phase A : 0.1 % FA in Water in Milli Q water, Mobile Phase B : Acetonitrile (100%).

[0211] Mobile phase gradient details: T = 0 min (90% A, 10% B) T = 7 min (10%A, 90% B) T = 9 min (0% A, 100% B) T = 14 min (0% A, 100% B) T = 14.01 min (90% A, 10% B) gradient to T= 17 min (90% A, 10% B)., Flow rate:- 1.0 ml / min, Run Time:- 17 min, UV Detection Method:- PDA.Method: I

[0212] Machine Details: - Water alliance e2695 with 2998 PDA detector (HP-04), Agilent Infinity II G6125C (HP-05), Column temperature: 25°C, Auto sampler temperature: 25°C, Mobile Phase A : 5mM Ammonium Bicarbonate in Water in Milli Q water, Mobile Phase B : Acetonitrile (100%).

[0213] Mobile phase gradient details: T = 0 min (90% A, 10% B) T = 7 min (10%A, 90% B) T = 9 min (0% A, 100% B) T = 14 min (0% A, 100% B) T = 14.01 min (90% A, 10% B) gradient to T= 17 min (90% A, 10% B)., Flow rate:- 1.0 ml / min, Run Time:- 17 min, UV Detection Method:- PDA.Purification methodsPreparative reverse-phase HPLC conditions

[0214] Preparative HPLC purification was performed by reverse phase HPLC using a Waters Fractionlynx preparative HPLC system (2525 pump, 2996 / 2998 UV / VIS detector, 2767 liquid handler) or an equivalent HPLC system such as a Gilson Trilution UV directed system. The Waters 2767 liquid handler acted as both auto-sampler and fraction collector. The columns used for the preparative purification of the compounds were a Waters Sunfire OBD Phenomenex Luna Phenyl Hexyl or Waters Xbridge Phenyl at 10 pm 19 x 150 mm or Waters CSH Phenyl Hexyl, 19 x 150, 5 pm column unless otherwise stated. Appropriate focused gradients were selected based on acetonitrile and MeOH solvent systems under either acidic or basic conditions. The modifiers used under acidic / basic conditions were formic acid or TFA (0.1 % V / V) and ammonium bicarbonate (10 mM) respectively. The purification was controlled by Waters Fractionlynx software through monitoring at 210-400 nm and triggered a threshold collection value at 260 nm and, when using the Fractionlynx, the presence of target molecular ion as observed under API conditions. Collected fractions were analysed by LCMS (Waters Acquity systems with Waters SQD).

[0215] Below is a list of methods and conditions used for preparative reverse phase HPLC purifications.Chiral Supercritical Fluid Chromatography (SFC) separation protocol

[0216] The enantiomeric separation of compounds was achieved by Supercritical Fluid Chromatography (SFC) using a Waters Thar Prepl 00 preparative SFC system (P200 CO2 pump, 2545 modifier pump, 2998 UV / VIS detector, 2767 liquid handler with Stacked Injection Module). The Waters 2767 liquid handler acted as both auto-sampler and fraction collector. Appropriate isocratic methods were selected based on MeOH, ethanol or isopropanol solvent systems under un-modified or basic conditions. The standard SFC method used was modifier, CO2, 100 mL / min, 120 Bar backpressure, 40°C column temperature. The modifier used under basic conditions was diethylamine (0.1 % V / V). The modifier used under acidic conditions was either formic acid (0.1 % V / V) or TFA (0.1 % V / V). The SFC purification was controlled by Waters Fractionlynx software through monitoring at 210-400 nm and triggered at a threshold collection value, typically 260 nm. Collected fractions were analysed by SFC (Waters / Thar SFC systems with Waters SQD). The fractions that contained the desired product were concentrated by vacuum centrifugation.

[0217] Below is a list of achiral SFC methods used for purificationPreparation of example compoundsExample _ 1 _ (E)- / V-(2-(3-Chloro-6-hvdroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-4-((2- fluoroethyl)(methyl)amino)but-2-enamideStep 1 : Ethyl (E)-4-((2-fluoroethyl)(methyl)amino)but-2-enoate

[0218] A suspension of potassium carbonate (587 mg, 4.2 mmol, 2.0 eq) and 2-fluoro- / V-methylethan- 1-amine hydrochloride (CAS 3832-36-8, 0.434 mg, 3.8 mmol, 1 .8 eq) in THF (5.0 mL) was treated dropwise with a solution of ethyl (E)-4-bromobut-2-enoate (CAS 37746-78-4, 0.29 mL, 2.1 mmol, 1.0 eq) in THF (1 mL). The reaction mixture was stirred at room temperature for 2 h. The reaction was quenched with water (20 mL) and extracted with EtOAc (3 x 30 mL). The combined organic phases were washed with brine (30 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure to afford the title compound (309 mg, Yield: 77%).

[0219] MS: ES+ 190.1 (M+1).

[0220] 1H NMR (400 MHz, CDCI3) 6, 1 .29 (t, J=7.1 Hz, 3H), 2.34 - 2.33 (m, 3H), 2.69 (t, J=4.9 Hz, 1 H), 2.76 (t, J=4.9 Hz, 1 H), 3.24 (dd, J=1 .5, 6.1 Hz, 2H), 4.20 (q, J=7.1 Hz, 2H), 4.49 (t, J=4.9 Hz, 1 H), 4.61 (t, J=5.0 Hz, 1 H), 6.02 - 5.97 (m, 1 H), 6.99 - 6.91 (m, 1 H).Step 2: (E)-4-((2-Fluoroethyl)(methyl)amino)but-2-enoic acid

[0221] To a solution of ethyl (E)-4-((2-fluoroethyl)(methyl)amino)but-2-enoate (Step 1) (300 mg, 1.6 mmol, 1.0 eq) in THF (6.5 mL) was added 2M aqueous sodium hydroxide (2.4 mL, 4.8 mmol, 3.0 eq) and the reaction mixture was stirred at room temperature for 16 h. The pH of the mixture was adjusted to pH 6 with 2N HCI and the solvent was removed under reduced pressure. The residue was stirred with 10% MeOH in DCM (50 mL) for 2 h then filtered. The filtrate was concentrated under reduced pressure to afford the title compound (256 mg, Yield: 100%).

[0222] 1H NMR (400 MHz, CDCI3) 6, 2.39 - 2.36 (m, 3H), 2.90 - 2.76 (m, 2H), 3.28 (d, J=6.0 Hz, 2H), 4.56 - 4.50 (m, 1 H), 4.65 (t, J=4.4 Hz, 1 H), 5.97 (d, J=15.6 Hz, 1 H), 6.84 - 6.73 (m, 1 H). Exchangeable protons not visible.

[0223] MS: ES+ 162.0 (M+1).Step 3: te / 7-Butyl (E)-4-(4-((2-fluoroethyl)(methyl)amino)but-2-enamido)isoindoline-2-carboxylate

[0224] To a stirred solution of (E)-4-((2-fluoroethyl)(methyl)amino)but-2-enoic acid (Step 2) (247 mg, 1 .5 mmol, 1.1 eq), HATU (604 mg, 1.6 mmol, 1.2 eq) and te / 7-butyl 4-aminoisoindoline-2-carboxylate (CAS 871013-98-8, 310 mg, 1 .3 mmol, 1 .0 eq) in DCM (6.0 mL) was added DIPEA (0.46 mL, 2.6 mmol, 2.0 eq) and the reaction mixture was stirred at room temperature for 2 h. The resulting mixture was diluted with DCM (30 mL) and washed with water (30 mL). The aqueous phase was re-extracted with DCM (2 x 30 mL) and the combined organic phases were washed with brine (30 mL), dried over MgSC , filtered, and concentrated under reduced pressure. The residue was purified by chromatography on silica eluting with 0 -10% MeOH in DCM to afford the title compound (541 mg, Yield: 83%).

[0225] MS:ES+ 378.1 (M+1).Step 4: (E)-4-((2-Fluoroethyl)(methyl)amino)- / V-(isoindolin-4-yl)but-2-enamide trifluoroacetate

[0226] To a solution of te / Y-butyl (E)-4-(4-((2-fluoroethyl)(methyl)amino)but-2-enamido)isoindoline-2- carboxylate (Step 3) (541 mg, 1 .10 mmol, 1 .0 eq) in DCM (5 mL) was added TFA (1 .7 mL, 22.2 mmol, 20.1 eq) and the reaction mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure to afford the title compound (432 mg, Yield: quantitative). The material was used in the next step without further purification.

[0227] MS:ES+ 278.1 (M+1).Step 5: (E)- / V-(2-(3-Chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-4-((2- fluoroethyl)(methyl)amino)but-2-enamide

[0228] To a stirred solution of (E)-4-[2-fluoroethyl(methyl)amino]- / V-isoindolin-4-yl-but-2-enamide;2,2,2- trifluoroacetic acid (Step 4) (432 mg, 1.10 mmol, 1 .0 eq), HOBt (169 mg, 1.10 mmol, 1.0 eq), 3-chloro-6- hydroxy-4-methoxy-2-methylbenzoic acid (Intermediate D) (239 mg, 1 .10 mmol, 1 .0 eq) and EDC (423 mg, 2.21 mmol, 2.0 eq) in DCM (9 mL) was added NMM (3.6 mL, 33.1 mmol, 30.0 eq). The reaction mixture was stirred at room temperature for 16 h under an inert atmosphere. The resulting mixture was diluted with DCM (40 mL) and washed with water (20 mL), brine (20 mL), filtered through a hydrophobic frit, and concentrated under reduced pressure. The crude residue was purified by achiral SFC (Method: SFC-1) to afford the title compound (17 mg, Yield: 3.1 %) as an off-white solid.

[0229] 1H NMR (400 MHz, DMSO) 6, 2.19 - 2.17 (m, 3H), 2.28 - 2.21 (m, 3H), 2.75 - 2.59 (m, 2H), 3.25- 3.16 (m, 2H), 3.82 - 3.81 (m, 3H), 4.64 - 4.40 (m, 4H), 4.83 - 4.76 (m, 2H), 6.37 (q, J=23.0 Hz, 1 H), 6.55- 6.54 (m, 1 H), 6.81 - 6.65 (m, 1 H), 7.17 - 7.02 (m, 1 H), 7.32 - 7.23 (m, 1 H), 7.74 - 7.64 (m, 1 H), 9.71 - 9.47 (m, 1 H), 10.06 (s,1 H).

[0230] 19F NMR (376 MHz, DMSO) -218 (s).

[0231] LCMS (LC-Method 1): 2.5 min, MS:ES+ 476.3 (M+1).

[0232] The compounds in the following table were prepared analogously to Example 1 , Step 5 from the indicated intermediates.Example _ 2: _ (E)- / V-(6-(3-Chloro-6-hvdroxy-4-methoxy-2-methylbenzoyl)-5,6,7,8-tetrahydro-1 ,6- naphthyridin-3-yl)-4-(dimethylamino)-A / -methylbut-2-enamide trifluoroacetateStep 1 : tert-Butyl 3-(methylamino)-7,8-dihydro-1 ,6-naphthyridine-6(5 / 7)-carboxylate

[0233] To a reaction tube charged with a degassed stirred solution of tert-butyl 3-bromo-7,8-dihydro- 5 / 7-1 ,6-naphthyridine-6-carboxylate (CAS 1184950-48-8) (1.5 g, 4.8 mmol, 1.0 eq) and sodium tert- butoxide (1.15 g, 12 mmol, 2.5 eq) in toluene (25 mL) was added 2M methylamine in THF (10 mL, 19mmol, 4.0 eq), tris(dibenzylideneacetone)dipalladium(0) (175 mg, 0.19 mmol, 0.04 eq) and BrettPhos (154 mg, 0.29 mmol, 0.06 eq). The reaction mixture was heated at 80°C under nitrogen for 18 h. The resulting mixture was allowed to cool, diluted with water (75 mL) and extracted with EtOAc (3 x 75mL). The combined organic extracts were washed with brine (100 mL), dried over MgSC , filtered, and concentrated under reduced pressure. The crude residue was purified by chromatography on silica eluting with 3:1 EtOAc: EtOH followed by 0-100% EtOAc in cyclohexane to afford the title compound as an orange oil (1 .2 g, Yield: 79%).

[0234] MS:ES+ 264.1 (M+1).

[0235] 1H NMR (400 MHz, DMSO) 6 1 .42 (s, 9H), 2.71 - 2.65 (m, 5H), 3.60 - 3.55 (m, 2H), 4.44 (s, 2H), 5.74 - 5.67 (m, 1 H), 6.65 (d, J = 3.0 Hz, 1 H), 7.77 (d, J = 2.5 Hz, 1 H).Step 2: (E)-4-(Dimethylamino)- / V-methyl- / V-(5,6,7,8-tetrahydro-1 ,6-naphthyridin-3-yl)but-2-enamide dihydrochloride

[0236] A solution of (E)-4-(dimethylamino)but-2-enoyl chloride hydrochloride (CAS 1055943-40-2) (839 mg, 4.6 mmol, 1.2 eq) in acetonitrile (12 mL) was added to te / Y-butyl 3-(methylamino)-7,8-dihydro-1 ,6- naphthyridine-6(5 / - / )-carboxylate (Step 1) (1 .15 g, 3.8 mmol, 1.0 eq) in acetonitrile (4.0 mL) under an inert atmosphere at 0°C. The reaction mixture was stirred under an inert atmosphere at 0°C for 1 h then concentrated under reduced pressure. The crude residue was dissolved in DCM (10 mL) and cooled to 0°C. 3M HCI in CPME (15 mL, 46 mmol, 12 eq) was added and the mixture stirred at 0°C for 1 h. The resulting mixture was concentrated under reduced pressure to afford the title compound (1.04 g, Yield: 79%). The material was used in the next step without further purification.

[0237] MS:ES+ 275.1 (M+1).Step 3: (E)- / V-(6-(3-Chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)-5,6,7,8-tetrahydro-1 ,6-naphthyridin-3- yl)-4-(dimethylamino)- / V-methylbut-2-enamide trifluoroacetate

[0238] To a stirred solution of 3-chloro-6-hydroxy-4-methoxy-2-methylbenzoic acid (Intermediate D, 325 mg, 1.50 mmol, 1.6 eq), HOBt (184 mg, 1.2 mmol, 1.2 eq) and EDC (460 mg, 2.4 mmol, 2.5 eq) in DCM (8.00 mL) and DMF (0.60 mL) was added NMM (1 .3 mL, 12 mmol, 12 eq). After stirring at room temperaturefor 5 mins, (E)-4-(dimethylamino)- / V-methyl- / V-(5,6,7,8-tetrahydro-1 ,6-naphthyridin-3-yl)but-2-enamide dihydrochloride (Step 2) (335 mg, 0.97 mmol, 1.0 eq) was added and stirring continued at room temperature under an inert atmosphere for 18 h. The resulting mixture was diluted with DCM (5 mL) and washed with water (5 mL). The organic phase was filtered through a hydrophobic frit and concentrated under reduced pressure. The crude residue was purified by reverse phase HPLC (Method: prep-LC-2) to afford the title compound as an off-white solid (70 mg, Yield: 15%).

[0239] 1H NMR (400 MHz, DMSO) 6 2.12 - 2.00 (m, 3H), 2.74 - 2.69 (m, 6H), 3.00 - 2.79 (m, 2H), 3.31 - 3.23 (m, 3H), 3.56 - 3.51 (m, 2H), 3.83 - 3.78 (m, 5H), 4.98 - 4.37 (m, 2H), 6.25 - 6.11 (m, 1 H), 6.51 (d, J = 10.5 Hz, 1 H), 6.71 - 6.61 (m, 1 H), 7.79 - 7.59 (m, 1 H), 8.37 - 8.31 (m, 1 H), 9.59 (s, 1 H), 10.04 - 9.97 (m, 1 H, TFA 1 eq).

[0240] LCMS (LC-Method 1): 2.6 min, MS:ES+ 473.3 (M+1).Example 3 (E)- / V-(5-Chloro-2-(5-chloro-2-hvdroxy-4-methoxybenzoyl)isoindolin-4-yl)-4-(dimethylamino)but-2-enamideStep 1 : te / Y-Butyl 4-amino-5-chloroisoindoline-2-carboxylate

[0241] To a stirred solution of fe / Y-butyl 4-aminoisoindoline-2-carboxylate (CAS 871013-98-8) (10.0 g, 0.042 mmol, 1 .0 eq.) in ACN (150 mL) was added NCS (5.71 g, 0.042 mmol, 1 .0 eq.) and the reaction mixture stirred at room temperature for 24 h. The resulting mixture was poured into water (400 mL), filtered, and the resultant solid washed with water (100 mL). The aqueous filtrate was extracted with EtOAc (5 x 100 mL) and the combined organic extracts were dried over Na2SC>4 and concentrated under reduced pressure. Crude material was purified by chromatography on silica (product eluted at 12.0% EtOAc in hexane) to yield fe / Y-butyl 4-amino-5-chloroisoindoline-2-carboxylate (1.7 g, Yield: 15%).

[0242] 1H NMR (DMSO-d6, 400 MHz): 6 1.45 (d, J= 5.6 Hz, 9H), 4.46 (dd, J = 21 .6 Hz, J = 11 .2 Hz, 4H), 5.37 (d, J = 8.8 Hz, 2H), 6.51 (t, J = 6.8 Hz, J= 14.0 Hz, 1 H), 7.12 (d, J = 8.0 Hz, 1 H).

[0243] LCMS (Method A): 2.357 min, MS: ES+ 213 (M-56).Step 2: te / Y-Butyl (E)-5-chloro-4-(4-(dimethylamino)but-2-enamido)isoindoline-2-carboxylatePyridine, POCI3, DCM, O-rt, 16h

[0244] The title compound was prepared from tert-butyl 4-amino-5-chloroisoindoline-2-carboxylate (Step 1) and (E)-4-(dimethylamino) but-2-enoic acid hydrochloride (CAS: 848133-35-7) analogously to Example 6, Step 1 .

[0245] 1H-NMR (DMSO-d6, 400 MHz): 6 1.45 (s, 9H), 2.19 (s, 6H), 3.07 (d, J = 5.6 Hz, 2H), 4.11 (s, br, 1 H, D2O exchangeable), 4.44 (d, J = 11 .2 Hz, 2H), 4.614 (d, J = 9.6Hz, 2H), 6.31 - 6.36 (m, 1 H), 6.73 - 6.78 (m, 1 H), 7.25 - 7.28 (m, 1 H), 7.46 (d, J = 8 Hz, 1 H), 9.90 (d, J = 14Hz, D2O exchangeable).

[0246] LCMS (Method: N): 1.723 min, MS ES+: 380.2, 382.2 (M, M+2).Step 3: (E)- / V-(5-Chloroisoindolin-4-yl)-4-(dimethylamino)but-2-enamide hydrochloride

[0247] The title compound was prepared from fert-butyl (E)-5-chloro-4-(4-(dimethylamino)but-2- enamido)isoindoline-2-carboxylate (Step 2) and 4M HCI in dioxane analogously to Example 6, Step 2.

[0248] 1H-NMR (DMSO-d6, 400 MHz): 6 2.77 and 2.78 (2 singlets, 6H), 3.95 (m, 2H), 3.17 (s, 2H), 4.35- 4.39 (m, br, 2H), 4.55 (s, br, 2H), 6.57 - 6.60 (m, 1 H), 6.83 - 6.90 (m, 1 H), 7.35 (d, J = 8 Hz, 1 H), 7.57 (d, J = 8 Hz, 1 H), 9.99 (s, br, 2H, D2O exchangeable), 10.85 (s, br, 1 H, D2O exchangeable).

[0249] LCMS (Method N): 0.305 min, MS: ES+ 280.2, 282.1 (M, M+2).Step 4: (E)- / V-(5-Chloro-2-(5-chloro-2-hydroxy-4-methoxybenzoyl)isoindolin-4-yl)-4-(dimethylamino)but-2- enamide

[0250] To a stirred solution of 5-chloro-2-hydroxy-4-methoxybenzoic acid (CAS: 1378866-39-7) (0.17 g, 0.839 mmol, 1 .0 eq.) in DMF (3.5 mL) at room temperature were added EDC.HCI (0.24 g, 1 .258 mmol, 1 .5 eq.) and HOAt (0.168 g, 0.839 mmol, 1.0 eq). After 10 mins, (E)- / V-(5-chloroisoindolin-4-yl)-4- (dimethylamino)but-2-enamide hydrochloride (Step 3) (0.212 g, 0.671 mmol, 0.8 eq) and / V-methyl morpholine (0.65 mL, 5.87 mmol, 7.0 eq) were added and the reaction mixture was stirred at room temperature for 50 mins. The resulting mixture was diluted with ice-cold water (50 mL) and extracted with ethyl acetate (2 x 100 mL). The combined organic extracts were washed with saturated brine solution (100 mL), dried over Na2SC>4, filtered, and concentrated under reduced pressure. The crude material was purified by reverse phase chromatography eluting with 45% MeCN in water. The product fractions were concentrated under reduced pressure and further purified by preparative HPLC (conditions shown below) followed by lyophilization of the pure fractions to afford (E)- / V-(5-chloro-2-(5-chloro-2-hydroxy-4- methoxybenzoyl)isoindolin-4-yl)-4-(dimethylamino)but-2-enamide as white solid (0.033 g, Yield: 10%).

[0251] HT1H NMR (DMSO-d6, 400 MHz, 349K): 5 2.21 (s, 6H), 3.09 (s, br, 2H), 3.86 (s, 3H), 4.67 (s, 2H), 4.82 (s, 2H), 6.29 - 6.33 (m, 1 H), 6.67 (s, 1 H), 6.75 - 6.79 (m, 1 H), 7.26 (d, J = 7.2 Hz, 1 H), 7.33 (s, 1 H), 7.45 (d, J = 8.0 Hz, 1 H), 9.65 (s, 1 H), 10.34 (bs, 1 H).

[0252] LCMS (Method N): 1.211 min, MS: ES+ 464.1 (M+1).

[0253] HPLC (Method H): 4.073 min.

[0254] Prep. HPLC purification method of analysis: Chromatographic separation and isolation used a Waters 2545 quaternary system with Waters 2489 UV Detector. Column: Waters X-BRIDGE PREP C18 (250 x 20 mm ID, 5pm); compounds eluted with, Mobile Phase A: 5 mM ammonium bicarbonate + 0.05 % ammonium hydroxide in water, Mobile Phase B: acetonitrile. Gradient T = 0.01 (70% A, 30% B) to T = 20.00 min (50% A, 50% B); T = 20.01 min (0% A, 100% B) gradient to T = 23.00 min (0% A, 100% B); T = 23.01 min (70% A, 30% B); gradient to T = 27.00 min (70% A, 30% B); Flow rate= 8ml / min; analysis time 27.00 min.Example 4: / V-(2-(3-Chloro-6-hvdroxy-4-methoxy-2-methylbenzoyl)-6-(2-(dimethylamino)ethoxy)isoindolin-4-yl)- / V-methylacrylamide

[0255] To a stirred solution 3-chloro-6-hydroxy-4-methoxy-2-methylbenzoic acid (Intermediate D) (0.2 g, 0.92 mmol, 1 eq) in DMF (2.0 mL) at room temperature, were added EDC.HCI (0.265 g, 1.3 mmol, 1.5 eq) and HOAt (0.125 g, 0.92 mmol, 1 eq) followed by / V-(6-(2-(dimethylamino)ethoxy)isoindolin-4-yl)- / V- methylacrylamide.TFA salt (Intermediate E) (0.56 g, 1 .39 mmol, 1.5 eq.) and NMM (0.46 g, 4.55 mmol, 5 eq.). The reaction mixture was stirred at room temperature for 1 h. The resulting mixture was poured into ice cold water (100 mL) and extracted into EtOAc (3 x 100 mL). The combined organic extracts were washed with ice-cold water (3 x 50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude material was purified by reverse phase chromatography followed by preparative HPLC purification (see below for conditions) to afford / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)-6- (2-(dimethylamino)ethoxy)isoindolin-4-yl)- / V-methylacrylamide as an off-white solid (0.028 g, Yield: 6%).

[0256] 1H NMR (DMSO, 400 MHz): 6 2.11 - 2.17 (m, 3H), 2.19 - 2.21 (m, 6H), 2.57 - 2.63 (m, 2H), 3.09 and 3.21 (2 singlets, 3H), 3.80 - 3.81 (m, 3H), 4.01 - 4.09 (m, 2H), 4.31 - 4.44 (m, 1 H), 4.55 - 4.58 (m, 1 H), 4.6 - 4.8 (m, 1 H), 4.82 (s, 1 H), 5.61 - 5.63 (m, 1 H), 6.06 - 6.11 (m, 1 H), 6.23 (d, J= 16.4 Hz, 1 H), 6.50 - 6.54 (m, 1 H), 6.87 (d, J= 6.8 Hz, 1 H), 6.93 - 7.07 (2 singlets, 1 H), 10.08 (s, 1 H).

[0257] LCMS (Method N): 1.188 min, MS: ES+ 488.2, 490.1 (M, M+2).

[0258] HPLC (Method H): 3.463 min.

[0259] Prep. HPLC purification and method of analysis: Chromatographic separation and isolation were conducted with a Waters 2545 quaternary system with Waters 2489 UV Detector. Column: Waters X- Bridge Prep C18 (150mm x 19mm 5pm); compounds eluted with Mobile Phase A: 0.05% ammonia + 5mMammonium bicarbonate in Merck water; Mobile Phase B: acetonitrile. Gradient T = 0.01 min (78% A, 22% B); gradient to T = 20.00 min (68% A, 32% B); T = 20.01 min (0% A, 100% B); gradient to T = 23.00 min (0% A, 100% B); T = 23.01 min (78% A, 22% B); T = 27.00 min (78% A, 22% B); Flow rate= 12 ml / min; analysis time 27 min.Example 5: (E)-4-(Dimethylamino)- / V-(2-(6-hvdroxy-4-methoxy-2,3-dimethylbenzoyl)isoindolin-4-yl)- / V- methylbut-2-enamideNMM, DMF, rt, 15min

[0260] The title compound was prepared from 6-hydroxy-4-methoxy-2,3-dimethylbenzoic acid (Intermediate F) and (E)-4-(dimethylamino)- / V-(isoindolin-4-yl)- / V-methylbut-2-enamide dihydrochloride (Intermediate S) analogously to Example 4.

[0261] 1H NMR (DMSO-d6, 400 MHz): 6 1 .91 - 2.10 (m, 9H), 2.20 (s, 3H), 2.85 - 3.01 (m, 2H), 3.08 and 3.21 (2 singlets, 3H), 3.74 (s, 3H), 4.11 - 4.88 (m, 4H), 5.68 - 5.81 (m, 1 H), 6.30 - 6.39 (m, 1 H), 6.60 - 6.71 (m, 1 H), 7.23 (d, br, J= 4.4 Hz, 1 H), 7.32 (d, J = 7.2 Hz, 1 H), 7.39 - 7.45 (m, 1 H), 9.47 - 9.51 (m, br, 1 H).

[0262] LCMS (Method N): 1.579 min, MS ES+: 438.3 (M+1).

[0263] HPLC (Method H): 4.20 min.

[0264] Prep. HPLC purification and method of analysis; Chromatographic separation and isolation were conducted with a Waters 2545 quaternary system with Waters 2489 UV Detector. Column: Waters X- Bridge C18 (250mm x 19mm x 5pm); compounds were eluted with: Mobile Phase A: 5mM ammonium bicarbonate +0.05% NH3 in Merck water; mobile Phase B: acetonitrile. Gradient of T = 0.01 min (68% A, 32% B); gradient to T = 22.00 min (68% A, 32% B); T = 22.01 min (0% A, 100% B), gradient to T = 26.00 min (0% A, 100% B); T = 26.01 min (68% A, 32% B); gradient to T = 30.00 min (68% A, 32% B); Flow rate= 11 ml / min; analysis time 30.00 min.Example 6: (E)- / V-(2-(3-Chloro-6-hvdroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)- / V-methyl-4-(methylamino)but-2-enamideStep 1 : te / Y-Butyl (E)-4-(4-(((benzyloxy)carbonyl)(methyl)amino)- / V-methylbut-2-enamido)isoindoline-2- carboxylateDCM,O°- rt, 2h

[0265] Performed in 2 parallel batches, each of 2.1 g scale: To a stirred solution of (E)-4- (((benzyloxy)carbonyl)(methyl)amino)but-2-enoic acid (Intermediate G) (4.20 g, 16.86 mmol, 1.0 eq) in DCM (84 mL) at room temperature was added fe / Y-butyl-4-(methylamino)isoindoline-2-carboxylate (Intermediate H) (2.51 g, 10.12 mmol, 0.6 eq). The mixture was cooled to 0°C, pyridine (5.33 g, 67.5 mmol, 4.0 eq) was added and stirring continued for 15 mins. POCh (2.34 mL, 25.30 mmol, 1.5 eq) was added dropwise at 0°C and the reaction mixture was allowed to warm to room temperature stirring for 1 h 45 mins. The resulting mixture was diluted with water (200 mL) and extracted with EtOAc (2 x 200 mL). The combined organic extracts were dried over Na2SC>4 and concentrated under reduced pressure. The crude material was purified by chromatography on silica (product eluted in 40% EtOAc: hexane) yielding tert- butyl (E)-4-(4-(((benzyloxy)carbonyl)(methyl)amino)- / V-methylbut-2-enamido)isoindoline-2-carboxylate as yellow liquid (3.0 g, Yield: 37%).

[0266] 1H NMR (DMSO-d6, 400 MHz): 6 1 .46 (s, 9H), 2.64 (m, br, 2H), 2.83 - 2.88 (m, 1 H), 3.17 (s, 3H), 3.87 (s, br, 2H), 4.35 (s, br, 1 H), 4.56 - 4.62 (m, 3H), 4.93 - 5.09 (m, 2H), 5.61 - 5.65 (m, 1 H), 6.63 (s, br, 1 H), 7.17 - 7.37 (m, 8H).

[0267] LCMS (Method N): 1.833 min, MS: ES+ 380.1 (M-100).Step 2: Benzyl-(E)-(4-(isoindolin-4-yl(methyl)amino)-4-oxobut-2-en-1 -yl) (methyl)carbamate hydrochloride

[0268] To a stirred solution of tert-butyl (E)-4-(4-(((benzyloxy)carbonyl) (methyl)amino)- / V-methylbut-2- enamido)isoindoline-2-carboxylate (Step 1) (3.0 g, 6.26 mmol, 1.0 eq) in DCM (30 mL) at room temperature, was added dropwise 4M HCI in dioxane (15 mL) at 0°C and the reaction mixture stirred for 1 h. The resulting mixture was diluted with DCM (10 mL) and concentrated under high vacuum yielding benzyl-(E)-(4-(isoindolin-4-yl(methyl)amino)-4-oxobut-2-en-1 -yl) (methyl)carbamate hydrochloride as a liquid (2.20 g, Yield: quantitative). The material was used in the next step without further purification.

[0269] LCMS (Method N): 1.216 min, MS: ES+ 380.2 (M-100).Step 3: Benzyl-(E)-(4-((2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl) isoindolin-4-yl) (methyl)amino)-4-oxobut-2-en-1-yl)(methyl)carbamateNMM, DMF, rt, 16h

[0270] The title compound was prepared from 3-chloro-6-hydroxy-4-methoxy-2-methylbenzoic acid (Intermediate D) and benzyl-(E)-(4-(isoindolin-4-yl(methyl)amino)-4-oxobut-2-en-1-yl) (methyl)carbamate hydrochloride (Step 2) analogously to Example 4.

[0271] 1H NMR (DMSO-d6, 400 MHz): 6 2.12 - 2.17 (m, 3H), 2.55 - 2.67 (m, 2H), 2.85 (s, 1 H), 2.89 - 3.38 (m, 4H), 3.74 - 4.09 (m, 5H), 4.41 - 4.70 (m, 3H), 4.75 - 5.20 (m, 2H), 5.62 (s, br, 1 H), 6.50 - 6.67 (m, 2H), 7.18 - 7.43 (m, 8H), 10.06 (bs, 1 H).

[0272] LCMS (Method N): 1.663 min, MS: ES+ 578.1 (M+1),Step 4: (E)- / V-(2-(3-Chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)- / V-methyl-4-(methylamino)but-2-enamide

[0273] To a stirred solution of benzyl (E)-(4-((2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl) isoindolin-4-yl) (methyl)amino)-4-oxobut-2-en-1-yl)(methyl)carbamate (Step 3) (0.45 g, 0.778 mmol, 1.0 eq) in hexafluoroisopropanol (13.5 mL) at room temperature was added portion wise anhydrous AlCh (0.518 g, 3.89 mmol, 5.0 eq) and the reaction mixture stirred for 16 h. The resulting mixture was diluted with aqueous sodium bicarbonate solution (50 mL) and extracted with EtOAc (3 x 50 mL). The combined organic extracts were dried over Na2SC>4 and concentrated under reduced pressure. The crude material was purified by chromatography on silica (product eluted in 12% MeOH: water) followed by preparative HPLC (see below for conditions) yielding (E)- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2- methylbenzoyl)isoindolin-4-yl)- / V-methyl-4-(methylamino)but-2-enamide as a white solid (0.015 g, Yield: 5%).

[0274] 1H NMR (DMSO-d6, 400 MHz, 349K): 6 2.16 - 2.21 (m, 6H), 3.10 (s, 3H), 3.23 (s, br, 2H), 3.82 (s, 3H), 4.45 - 4.89 (m, 4H), 5.83 (s, br, 1 H), 6.47 - 6.76 (m, 2H), 7.19 (d, J= 7.6 Hz, 1 H), 7.28 (d, J= 8.0 Hz, 1 H), 7.37 - 7.44 (m, 1 H), 8.26 (s, 1 H, formate salt).

[0275] LCMS (Method N): 1.192 min, MS: ES+ 444.2 (M+1).

[0276] HPLC (Method I): 5.28 min.

[0277] Prep. HPLC purification and method of analysis: Chromatographic separation and isolation were conducted with a Waters 2545 quarternary system with Waters 2489 UV Detector; column: Sunfire Prep C18 (250 x 19 mm, 5um); compounds eluted with Mobile Phase A: 0.05% formic acid in Merck water, Mobile Phase B: Acetonitrile. Gradient: T = 0.00 min (95% A, 5% B); gradient to T = 18.00 min (77% A,23% B); T = 18.01 min (0% A, 100%B); gradient to T = 20.00 min (0% A, 100% B); T = 20.01 min (95% A, 5% B); T = 22.00 min (95% A, 5% B); Flow rate= 15 ml / min; analysis time 22 min.Example 7: (E)-4-(Dimethylamino)- / V-(2-(2-hvdroxy-5-isopropyl-4-methoxybenzoyl)-3-methyl-1 ,2,3,4- tetrahvdroisoquinolin-7-yl)- / V-methylbut-2-enamideStep 1 : (E)-4-(Dimethylamino)- / V-(2-(2-hydroxy-5-isopropyl-4-methoxybenzoyl)-3-methyl-1 ,2,3,4- tetrahydroisoquinolin-7-yl)- / V-methylbut-2-enamide

[0278] To a stirred solution 2-hydroxy-5-isopropyl-4-methoxybenzoic acid (Intermediate I) (0.09 g, 0.428 mmol, 1 eq) in DMF (1 .8 mL) at room temperature were added EDC.HCI (0.123 g, 0.642 mmol, 1.5 eq) and HOAt (0.058 g, 0.428 mmol, 1 eq) and the mixture stirred for 15 mins. (E)-4-(dimethylamino)- / V-methyl- / V-(3-methyl-1 ,2,3,4-tetrahydroisoquinolin-7-yl)but-2-enamide hydrochloride (Intermediate J) (0.166 g, 0.51 mmol, 1.2 eq) and / V-methyl morpholine (0.216 g, 2.14 mmol, 5.0 eq.) were added and stirring continued at room temperature for 4 h. The resulting mixture was poured into cold water (100 mL) and extracted with ethyl acetate (3 x 50 mL). The combined organic extracts were dried over Na2SC>4, filtered, and concentrated under reduced pressure. The crude material was purified by preparative HPLC (see conditions below) yielding (E)-4-(dimethylamino)- / V-(2-(2-hydroxy-5-isopropyl-4-methoxybenzoyl)-3- methyl-1 ,2,3,4-tetrahydroisoquinolin-7-yl)- / V-methylbut-2-enamide as a white solid (0.040 g, Yield: 22%).

[0279] 1H NMR (DMSO-d6, 400 MHz): 6 1.06 - 1 .24 (m, 9H), 2.02 (s, 6H), 2.63 (d, J= 16.4 Hz, 1 H), 2.87 (d, J= 5.6 Hz, 2H), 3.05 - 3.21 (m, 2H), 3.34 (s, 3H), 3.77 (s, 3H), 4.28 (d, J= 14 Hz, 1 H), 5.02 (s, br,1 H), 5.87 (d, J= 13.6 Hz, 1 H), 6.47 (s, 1 H), 6.56 - 6.63 (m, 1 H), 6.92 (s, 1 H), 7.07 - 7.23 (m, 3H), 9.71 (s, 1 H).

[0280] LCMS (Method N): 1.805 min, MS: ES+ 480.31 (M+1).

[0281] HPLC (Method H): 4.71 min, 220 nm.

[0282] Prep. HPLC purification and method of analysis: Chromatographic separation and isolation were conducted with a Waters 2545 binary pump with Waters 2489 UV detector and Waters Acquity QDA detector. Column YMC Triart C18 (250 x 20 mm ID, 5pm); compounds eluted with Mobile Phase A: 5mM ammonium bicarbonate +0.05% NH3 in Merck water, Mobile Phase B: Acetonitrile. Gradient T = 0.01 min (49% A, 51 % B) to T = 24.00 min (49% A, 51 % B); T = 24.01 min (00% A, 100% B) gradient to T = 27.00 min (0% A, 100% B); T = 27.01 min (49% A, 51 % B); gradient to T = 34.00 min (49% A, 51 % B); Flow rate= 16 ml / min; analysis time 34 min.Example 8: (E)-4-Amino- / V-(2-(3-chloro-6-hvdroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)- / V- methylbut-2-enamide hydrochlorideStep T te / Y-Butyl (E)-(4-((2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4- yl)(methyl)amino)-4-oxobut-2-en-1-yl)carbamate

[0283] To a stirred solution of (E)-4-((f-butoxycarbonyl)amino)but-2-enoic acid (Intermediate L) (0.2 g, 0.99 mmol, 1 .0 eq) in DCM (6 mL) at room temperature were added (3-chloro-6-hydroxy-4-methoxy-2- methylphenyl)(4-(methylamino)isoindolin-2-yl)methanone (Intermediate K) (0.206 g, 0.596 mmol, 0.6 eq) and pyridine (0.32 mL, 3.97 mmol, 4.0 eq). The reaction mixture was stirred at room temperature for 42 h then cooled to 0°C. POCh (0.13 mL, 1 .49 mmol, 1 .5 eq) was added dropwise and the mixture was allowed to warm to room temperature and stirred for 5 h. The resulting mixture was diluted with water (50 mL) and extracted with ethyl acetate (3 x 50 mL). The combined organic extracts were dried over Na2SC>4, filtered, and concentrated under reduced pressure. The crude material was purified by chromatography on silica (100% ethyl acetate) followed by preparative HPLC (see below for conditions) yielding te / Y-butyl (E)-(4-((2- (3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)(methyl)amino)-4-oxobut-2-en-1- yl)carbamate as an off-white solid (0.026 g, Yield: 8.51 %).

[0284] 1H NMR (DMSO-d6, 400 MHz): 6 1.23 - 1.29 (m, 9H), 2.17 (s, 3H), 3.06 (d, J= 6 Hz, 1 H), 3.19 (s, 2H), 3.54 - 3.58 (m, 2H), 3.81 (s, 3H), 4.41 - 4.85 (m, 4H), 5.58 - 5.70 (m, 1 H), 6.52 - 6.75 (m, 2H), 6.98 (bs, 1 H), 7.16 - 7.44 (m, 3H), 10.10 (s, 1 H).

[0285] LCMS (Method N): 1.528 min, MS: ES+ 474.1 (M-56).

[0286] HPLC (Method H): 6.41 min.

[0287] Prep. HPLC purification and method of analysis: Chromatographic separation and isolation were conducted with a Waters 2545 binary pump with Waters 2489 UV detector. Column REDISEP GOLD C18 100g 50pm; compounds eluted with: Mobile Phase A: 5mM ammonium bicarbonate +0.05%NH3 in Merck water, Mobile Phase B: Acetonitrile. Gradient of T = 0.00 min (100% A, 0% B) to T = 2.00 min (100% A, 0% B); T = 25.00 min (70% A, 30% B) gradient to T = 25.01 min (0% A, 100% B); T = 28.00 min (0% A, 100% B) gradient to T = 30.00 min (100% A, 0% B); Flow rate= 70 ml / min; analysis time 30 min.Step 2: (E)-4-Amino- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)- / V-methylbut-2- enamide hydrochloride

[0288] To a stirred solution of te / Y-butyl (E)-(4-((2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl) isoindolin-4-yl) (methyl)amino)-4-oxobut-2-en-1-yl) carbamate (Step 1) (0.02 g, 0.0377 mmol, 1.0 eq) in DCM (0.2 mL) cooled to 0°C was added dropwise 4M HCI in dioxane (0.1 mL). The reaction mixture was allowed to warm to room temperature stirring for 2 h. The resulting mixture was concentrated under high vacuum and the crude material was co-d istilled with n-pentane (2 x 5 mL) and n-hexane (2 x 5 mL) followedby lyophilization to yield (E)-4-amino- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4- yl)- / V-methylbut-2-enamide hydrochloride as a white solid (0.013 g, Yield: quantitative).

[0289] HT1H NMR (DMSO-d6, 400 MHz, 348K): 6 2.21 (s, 3H), 3.27 (s, br, 3H), 3.55 (s, br, 2H), 3.83 (s, br, b 3H), 4.50 (d, J= 14.8 Hz, 1 H), 4.66 (d, J= 15.2 Hz, 2H), 6.0 (s, br, 1 H), 6.62 - 6.82 (m, 2H), 7.18 - 7.46 (m, 3H), 9.88 (s, 1 H).

[0290] LCMS (Method N): 1.175 min, MS: ES+ 430.1 (M+1).

[0291] HPLC (Method H): 3.98 min.Example 9: (E)- / V-(2-(3-Chloro-6-hvdroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)- / V-methyl-4-(pyrrolidin-1-yl)but-2-enamidemicrowave

[0292] To a stirred solution of (3-chloro-6-hydroxy-4-methoxy-2-methylphenyl)(4- (methylamino)isoindolin-2-yl)methanone (Intermediate K) (0.15 g, 0.43 mmol, 1.0 eq.) in DMF (1.5mL) at room temperature were added DCC (0.178 g, 0.86 mmol, 2.0 eq.) and (E)-4-(pyrrolidin-1-yl)but-2-enoic acid hydrochloride (CAS: 848133-09-5) (0.082 g, 0.43 mmol, 1.0 eq.). The reaction mixture was heated using microwave irradiation at 150°C for 15 mins. The resulting mixture was poured into ice cold water (30 mL) and filtered through a Buchner funnel. The filtrate was concentrated under reduced pressure and the crude material was purified by reverse phase chromatography (product eluted at 48% water in acetonitrile) followed by preparative HPLC (conditions shown below) to yield (E)- / V-(2-(3-chloro-6-hydroxy-4-methoxy- 2-methylbenzoyl)isoindolin-4-yl)- / V-methyl-4-(pyrrolidin-1-yl) but-2-enamide as a white solid (0.015 g, Yield: 4.8%).

[0293] 1H NMR (DMSO-d6, 400 MHz): 6 1.56 - 1.63 (m, 3H), 2.08 - 2.20 (m, 3H), 2.25 (s, br, 3H), 2.38 (s, br, 2H), 2.98 - 2.99 (m, 1 H), 3.08 - 3.20 (m, 2H), 3.40 (s, 2H), 3.76 (s, br, 3H), 4.41 - 4.57 (m, 2H), 4.65 - 4.85 (m, 2H), 5.70 - 5.80 (m, 1 H), 6.46 - 6.50 (m, 1 H), 6.63 - 6.74 (m, 1 H), 7.22 - 7.24 (m, 1 H), 7.29 -7.41 (m,1 H), 7.43 - 7.45 (m,1 H).

[0294] LCMS (Method N): 1.224 min, MS: ES+ 484.2, 486.1 (M, M+2).

[0295] HPLC (Method I): 6.01 min.

[0296] Prep. HPLC purification method of analysis: Chromatographic separation and isolation were conducted with Waters 2545 quaternary system with Waters 2489 UV Detector; column Waters X-BRIDGE C18 (250 x 20 mm ID, 5pm); compounds eluted with: Mobile Phase A: 0.05% NH3 in Merck water, Mobile Phase B: Acetonitrile. Gradient T = 0.00 (62% A, 38% B) to T = 19.00 min (47% A, 67% B); T = 19.01 min (0% A, 100% B) gradient to T = 21 .00 min (0% A, 100% B); T = 21 .01 min (62% A, 38% B); gradient to T = 25.00 min (62% A, 38% B); Flow rate= 7 ml / min; analysis time 25.00 min.Example 10: (E)- / V-(2-(3-Chloro-6-hvdroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-4-(pyrrolidin-1 - yl)but-2-enamide

[0297] To a stirred solution of (E)-4-(pyrrolidin-1-yl)but-2-enoic acid hydrochloride (CAS 848133-09-5; 0.19 g, 0.99 mmol, 1.0 eq.) and (4-aminoisoindolin-2-yl)(3-chloro-6-hydroxy-4-methoxy-2- methylphenyl)methanone (Intermediate KA) (0.13 g, 0.39 mmol, 0.4 eq.) in DCM (3.8 mL) at room temperature was added pyridine (1.9 mL). The reaction mixture was stirred at room temperature for 2 h then cooled to 0°C. POCh (0.13 mL, 1 .49 mmol, 1 .5 eq.) was added dropwise to the reaction mixture and the mixture allowed to warm to room temperature whilst stirring for 4 h. The resulting mixture was poured into water (100 mL) and extracted with DCM (100 mL). Both the organic and aqueous phases were concentrated under reduced pressure and the crude material was purified by reverse phase column chromatography (product eluted in 35% acetonitrile in water). The pure fractions were combined and concentrated under reduced pressure and further purified by preparative HPLC (see conditions below). The product fractions were lyophilized to yield (E)- / V-(2-(3-chloro-6-hydroxy-4-methoxy-2- methylbenzoyl)isoindolin-4-yl)-4-(pyrrolidin-1-yl) but-2-enamide an off-white solid (0.008 g, Yield: 2%).

[0298] 1H NMR (DMSO-d6, 400 MHz): 6 1 .70 (d, J = 16.8 Hz, 4H), 2.17 (s, 3H), 2.43 (s, 3H), 3.18 - 3.26 (m, 3H), 3.79 (s, 3H), 4.40 - 4.46 (m, 1 H), 4.57 (d, J= 23.2 Hz, 1 H), 4.77 - 4.81 (m, 2H), 6.28 - 6.46 (m, 1 H), 6.53 (s, 1 H), 6.71 - 6.83 (m, 1 H), 7.02 - 7.16 (m, 1 H), 7.23 - 7.31 (m, 1 H), 7.66 - 7.73 (m, 1 H), 9.46 - 9.69 (m, 1 H), 10.24 (bs, 1 H).

[0299] LCMS (Method: N): 1 .235 min, MS ES+: 470.1 , 472.2 (M, M+2).

[0300] HPLC (Method: I): 5.89 min.

[0301] Prep. HPLC purification and method of analysis: Chromatographic separation and isolation were conducted on a Waters 2545 quaternary system with Waters 2489 UV Detector. Column X-BRIDGE C18(150 x 19 mm ID, 5pm); compounds eluted with: Mobile Phase A: 5 mM Ammonium bicarbonate + 0.05% NH3 in water. Mobile Phase B:Acetonitrile. Gradient T = 0.00 min (85% A, 15% B) to T = 2.00 min (72% A, 28% B); T = 23.00 min (62% A, 38% B), gradient to T = 23.01 min (0% A, 100% B); T = 26.00 min (00% A, 100% B); gradient to T = 26.01 min (85% A, 15% B), gradient to T = 32.00 min (85% A, 15% B); Flow rate= 8.00 ml / min; analysis time 32.00 min.Example 11 : (E)- / V-(2-(3-Chloro-6-hvdroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)- / V-methyl-3-(1- methylpyrrolidin-2-yl)acrylamideStep 1 : Ethyl-(E)-3-(1-methylpyrrolidin-2-yl)acrylate

[0302] A stirred solution of fe / Y-butyl-(E)-2-(3-ethoxy-3-oxoprop-1-en-1-yl)pyrrolidine-1 -carboxylate (Intermediate M) (2.5 g, 9.29 mmol, 1.0 eq) in formic acid (30.35, 659.85 mmol, 71.0 eq.) was stirred at room temperature for 15 mins and then treated with paraformaldehyde (1 .39 g, 46.46 mmol, 5.0 eq.). The reaction mixture was stirred at 90°C for 4 h. The resulting mixture was poured into water (10 mL) and acidified with 1 M HCI solution (1 mL). The mixture was extracted with EtOAc (20 mL). The aqueous portion was basified with solid K2CO3 and extracted with EtOAc (3 x 100 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude material was purified by C18 reverse phase chromatography (product eluted in 27% acetonitrile in water). The pure fractions were combined and concentrated under reduced pressure to yield ethyl (E)-3-(1-methylpyrrolidin-2-yl) acrylate as a brown solid (0.3 g, 18% yield) which was used directly in the next step.

[0303] LCMS (Method: N): 0.861 min, MS ES+: 184.1 (M+1).Step 2: (E)-3-(1-Methylpyrrolidin-2-yl)acrylic acid

[0304] To a stirred solution of (E)-3-(1-methylpyrrolidin-2-yl)acrylate (Step 1) (0.3 g, 1 .63 mmol, 1 .0 eq.) in isopropylalcohol (3 mL) and water (3 mL) was added NaOH (0.523 g, 13.10 mmol, 8.0 eq.) and the reaction mixture stirred at room temperature for 16 h. The resulting mixture was poured into water (20 mL) and washed with EtOAc (50 mL). The aqueous portion was acidified with a saturated solution of KHSO4 (6 mL) and concentrated under reduced pressure. The crude material was diluted with 50% isopropylalcohol in CHCh (100 mL) and stirred at room temperature for 1 h. The mixture was filtered and the filtrate was concentrated under reduced pressure to yield (E)-3-(1-methylpyrrolidin-2-yl)acrylic acid as a brown gummy solid (0.22 g, 86% yield). The material was used in the next step without further purification.

[0305] 1H NMR (DMSO-d6, 400 MHz, D2O exchange): 6 1.82 - 1 .88 (m, 1 H), 1 .91 - 2.06 (m, 2H), 2.25 - 2.28 (m, 1 H), 2.73 and 2.74 (2 singlets, 3H), 3.08 - 3.17 (m, 2H), 3.92 - 4.04 (m, 1 H), 6.20 (d, J = 15.6Hz 2H), 6.82 dd, J = 15.6 Hz, J = 8.8Hz).

[0306] LCMS (Method: N): 0.295 min, MS ES+: 156.03 (M+1).Step 3: (E)- / V-(2-(3-Chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)- / V-methyl-3-(1- methylpyrrolidin-2-yl)acrylamide, m n, Microwave

[0307] The title compound was prepared from (E)-3-(1-methylpyrrolidin-2-yl) acrylic acid (Step 2) and (3-chloro-6-hydroxy-4-methoxy-2-methylphenyl)(4-(methylamino)isoindolin-2-yl)methanone (Intermediate K) analogously to Example 9.

[0308] 1H NMR (DMSO-d6, 400 MHz, D2O exchange): 6 1.40 - 1.44 (m, 1 H), 1.62 (bs, 2H), 1.82 - 2.17 (m, 9H), 2.91 (bs, 1 H), 3.08 and 3.21 (2 singlets, 3H), 3.85 (s, 3H), 4.41 - 4.89 (m, 4H), 5.63 - 5.78 (m, 1 H), 6.42 - 6.54 (m, 2H), 7.25 - 7.32 (m, 2H), 7.41 - 7.45 (m, 1 H).

[0309] LCMS (Method: N): 1.190 min, MS ES+: 484.2 (M).

[0310] HPLC (Method: I): 5.241 min.

[0311] Prep. HPLC purification and method of analysis: Chromatographic separation and isolation were conducted on a Waters 2545 quaternary system with Waters 2489 UV Detector. Column REDISEP RF C18 100g 50|jm. Compounds were eluted with: Mobile Phase A: 5 mM ammonium bicarbonate + 0.05% ammonia in Merck water. Mobile Phase B:Acetonitrile. Gradient T = 0.00 min (90% A, 10% B) to T = 25.00 min (40% A, 60% B); T = 25.01 min (00% A, 100% B), gradient to T = 28.00 min (0% A, 100% B); T = 28.01 min (90% A, 10% B); gradient to T = 32.00 min (90% A, 10% B); Flow rate= 70 ml / min; analysis time 32.00 min.Example 12: (E)- / V-(2-(3-Chloro-6-hvdroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)- / V-methyl-3-(pyrrolidin-2-yl)acrylamide hydrochlorideStep 1 : (E)-3-(1-(fe / Y-Butoxy carbonyl) pyrrolidin-2-yl) acrylic acid

[0312] The title compound was prepared from fe / Y-butyl-(E)-2-(3-ethoxy-3-oxoprop-1-en-1- yl)pyrrolidine-1 -carboxylate (Intermediate M) in IPA: H2O (1 :1) and NaOH analogously to Example 11 , Step 2.

[0313] 1H NMR (DMSO, 400 MHz, D2O exchange): 6 1 .34 - 1.40 (m, 9H), 1.69 - 1.78 (m, 3H), 1.91 - 2.04 (m, 1 H), 3.25 -3.29 (m, 2H), 4.29 - 4.35 (m, 1 H), 5.68 (d, J= 14.8 Hz, 1 H), 6.67 - 6.71 (m, 1 H).Step 2: te / Y-Butyl (E)-2-(3-((2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4- yl)(methyl)amino)-3-oxoprop-1 -en-1 -y I) py rro I id ine- 1 -carboxylate, ,

[0314] The title compound was prepared from (E)-3-(1-(fe / Y-butoxy carbonyl) pyrrolidin-2-yl) acrylic acid (Step 1) and (3-chloro-6-hydroxy-4-methoxy-2-methylphenyl)(4-(methylamino)isoindolin-2-yl)methanone (Intermediate K) analogously to Example 8, Step 1 .

[0315] 1H NMR (DMSO, 400 MHz, D2O exchange): 5 1.19 (s, 9H), 1.34 - 1.40 (m, 1 H), 1 .54 - 1.71 (m, 3H), 1.91 - 1.92 (m, 1 H), 2.17 (s, 3H), 2.92 - 3.13 (m, 3H), 3.21 (s, 2H), 3.81 (s, 3H), 4.11 - 4.87 (m, 4H), 5.49 - 5.58 (m, 1 H), 6.52 - 6.66 (m, 2H), 7.20 - 7.45 (m, 3H ) 9.59 - 9.60 (m, 1 H).

[0316] LCMS (Method N): 1.662 min, ES+ 514.1 (M-56).

[0317] HPLC (Method I): 6.79 min.

[0318] Prep. HPLC purification method of analysis: Chromatographic separation and isolation were conducted on a Waters 2545 quaternary system with Waters 2489 UV Detector. Column X-BRIDGE C18 (250 x 20 mm ID, 5pm). Compounds eluted with Mobile Phase A: 5mM ammonium bicarbonate + 0.05%NH3 in Merck water. Mobile Phase B:Acetonitrile. Gradient T = 2.0 min (56% A, 44% B) to T = 30.00 min (56% A, 44% B); T = 30.01 min (0% A, 100% B) gradient to T = 33.00 min (0% A, 100% B); T = 33.01 min (56% A, 44% B); T = 35.00 min (56% A, 44% B); Flow rate= 8 ml / min; analysis time 35 min.Step 3: (E)- / V-(2-(3-Chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)- / V-methyl-3-(pyrrolidin-2-yl)acrylamide hydrochloride

[0319] The title compound was prepared from te / Y-butyl (E)-2-(3-((2-(3-chloro-6-hydroxy-4-methoxy-2- methylbenzoyl) isoindolin-4-yl)(methyl)amino)-3-oxoprop-1-en-1-yl)pyrrolidine-1 -carboxylate (Step 2) and 4M HCI in dioxane analogously to Example 8, Step 2.

[0320] HT1H NMR (DMSO, 400 MHz, 348K): 5 1.66 - 1.67 (m, 1 H), 1.89 - 1.92 (m, 2H), 2.06 (s, 1 H), 2.20 (s, 3H), 3.27 (s, 3H), 3.83 (s, 3H), 4.09 - 4.10 (m, 1 H), 4.2 - 4.4 (m, 1 H), 4.48 - 4.52 (m, 1 H), 4.65 - 4.69 (m, 1 H), 4.90 (s, 1 H), 6.03 (s, br, 1 H), 6.63 (s, 1 H), 6.74 - 6.87 (m, 1 H), 7.21 (d, J= 7.6 Hz, 1 H), 7.32 (s, 1 H), 7.45 (s, 1 H), 8.98 - 8.99 (m, 1 H), 9.46 - 9.47 (m, 1 H), 9.90 - 9.91 (m, 1 H).

[0321] LCMS (Method N): 1.211 min, MS: ES+ 470.2 (M).

[0322] HPLC (Method H): 4.20 min.

[0323] Prep. HPLC purification and method of analysis: Chromatographic separation and isolation were conducted on a Waters 2545 quaternary system with Waters 2489 UV Detector. Column: Shim-Pack GIST C18 (250mm x 20mm x 5pm). Compounds eluted with Mobile Phase A: 0.05% hydrochloric acid in Merckwater. Mobile Phase B:Acetonitrile. Gradient T = 0.00 min (85% A, 15% B); to T = 18.00 min (65% A, 35% B); T = 18.01 min (0% A, 100% B); gradient to T = 20.00 min (0% A, 100% B); T = 20.01 min (85% A, 15% B); T = 25.00 min (85% A, 18% B); Flow rate= 18 ml / min; analysis time 25 min.Example 13: (E)- / V-(5-Chloro-2-(5-cvclopropyl-2-hvdroxy-4-methoxybenzoyl)isoindolin-4-yl)-4-(dimethylamino)but-2-enamide

[0324] The title compound was prepared from 5-cyclopropyl-2-hydroxy-4-methoxybenzoic acid (Intermediate N) and (E)- / V-(5-chloroisoindolin-4-yl)-4-(dimethylamino)but-2-enamide hydrochloride (Example 3, Step 3) analogously to Example 4.

[0325] 1H NMR (DMSO-d6, 400 MHz, D2O exchange): 6 0.54 (s, br, 2H), 0.79 (d, J= 7.6 Hz, 2H), 1 .92 - 1.97 (m, 1 H), 2.18 (d, J= 14.4 Hz, 6H), 3.06 (d, J= 16 Hz, 2H), 3.79 (s, 3H), 4.64 (s, 2H), 4.79 (d, J= 46 Hz, 2H), 6.25 - 6.39 (m, 1 H), 6.49 (m, 1 H), 6.69 - 6.78 (m, 2H), 7.24 - 7.34 (m, 1 H), 7.44 - 7.48 (m, 1 H).

[0326] LCMS (Method: N): 1.295 min, MS ES+: 470.2, 472.2 (M, M+2).

[0327] HPLC (Method: I): 6.752 min.

[0328] Prep. HPLC purification method of analysis: Chromatographic separation and isolation were conducted using a Waters 2545 binary pump with Waters 2489 UV detector and Acquity QDA detector; column Shim-Pack GIST C18 (250mm x 20mm x 5pm). Compounds eluted with: Mobile Phase A: 0.05% formic acid in water. Mobile Phase B:Acetonitrile. Gradient T = 0.00 min (80% A, 20% B) to T = 20.00 min (72% A, 28% B); T = 28.00 min (72% A, 28% B), gradient to T = 28.01 min (3% A, 97% B); T = 31 .00 min (3% A, 97% B); gradient to T = 31 .01 min (80% A, 20% B); T = 36.00 min (80% A, 20% B); Flow rate= 15 ml / min; analysis time 36.00.Example 14: (E)- / V-(2-(3-Chloro-6-hvdroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-4-(isopropylamino)but-2-enamideStep 1 : Ethyl (E)-4-(isopropylamino)but-2-enoate

[0329] To a suspension of potassium carbonate (1.43 g, 10.4 mmol, 2.0 eq) and isopropylamine (1.3 mL, 15.5 mmol, 3.0 eq) in THF (9.0 mL) was added dropwise a solution of ethyl 4-bromocrotonate (CAS: 37746-78-4, 0.71 mL, 5.18 mmol, 1.0 eq) in THF (3.0 mL) and the reaction mixture was stirred at room temperature for 2 h. The reaction was quenched with water (20 mL) and extracted with ethyl acetate (3 x20 mL). The combined organic extracts were washed with brine (20 mL), filtered through hydrophobic filter paper and concentrated under reduced pressure to afford the title compound (633 mg, Yield: 71 %).

[0330] 1H NMR (400 MHz, CDCb) 6 1 .07 (d, J=6.3 Hz, 6H), 1 .28 (t, J=7.1 Hz, 3H), 2.88 - 2.81 (m, 1 H), 3.41 (dd, J=1 .8, 5.6 Hz, 2H), 4.22 - 4.16 (m, 2H), 6.0 - 5.94 (m, 1 H), 7.04 - 6.97 (m, 1 H).Step 2: Ethyl (E)-4-[fe / Y-butoxycarbonyl(isopropyl)amino]but-2-enoate

[0331] A solution of ethyl (E)-4-(isopropylamino)but-2-enoate (Stepl) (633 mg, 3.70 mmol, 1.0 eq) and sodium carbonate (784 mg, 7.39 mmol, 2.0 eq) in THF (1 .85 mL) and water (1 .85 mL) at room temperature was treated with di-te / Y-buty I dicarbonate (1 .7 mL, 7.39 mmol, 2.0 eq) and the reaction mixture was stirred under an inert atmosphere for 2 h. The resulting mixture was diluted with DCM (40 mL), washed with water (2 x 20 mL) and brine (40 mL). The combined organic extracts were dried over MgSCU, filtered, and concentrated under reduced pressure. The crude residue was purified by chromatography on silica, eluting with a gradient of 0 - 50 % ethyl acetate in cyclohexane to afford the title compound (595 mg, Yield: 59%).

[0332] 1H NMR (400 MHz, CDCb) 6 1 .13 - 1 .09 (m, 6H), 1 .31 - 1 .26 (m, 3H), 1 .45 (s, 9H), 1 .59 - 1 .51 (m, 1 H), 3.95 - 3.76 (m, 2H), 4.23 - 4.15 (m, 2H), 5.90 - 5.84 (m, 1 H), 6.93 - 6.83 (m, 1 H).Step 3: (E)-4-[fe / Y-butoxycarbonyl(cyclopropyl)amino]but-2-enoic acid

[0333] To a solution of ethyl (E)-4-[fe / Y-butoxycarbonyl(isopropyl)amino]but-2-enoate (Step 2) (595 mg, 2.19 mmol, 1 .0 eq) in THF (9.0 mL) was added 2M sodium hydroxide (3.3 mL, 6.58 mmol, 3.0 eq) and the reaction mixture was stirred at room temperature for 2 h. Additional 2M sodium hydroxide (3.3 mL, 6.58 mmol, 3.0 eq) was added and the mixture was stirred at 40°C for 48 h. Additional 2 M sodium hydroxide (10 mL, 20.0 mmol, 9.12 eq) and THF (10.0 mL) was added and the mixture stirred at 80°C for a further 4 h. The mixture was allowed to cool to room temperature. The reaction mixture was adjusted to pH = 6 with 2N HCI, then concentrated under reduced pressure. The crude residue was stirred in 10% methanol in DCM (50 mL) for 2 h. The organics were dried over MgSCU, filtered, and concentrated under reduced pressure to afford the title compound (672 mg, Yield: 100%) which was used without further purification.Step 4: te / Y-Butyl 4-[[(E)-4-[tert-butoxycarbonyl(isopropyl)amino]but-2-enoyl]amino]isoindoline-2- carboxylate

[0334] (E)-4-[fe / Y-butoxycarbonyl(isopropyl)amino]but-2-enoic acid (Step 3) (311 mg, 1.28 mmol, 1.16 eq), HATU (504 mg, 1.33 mmol, 1.20 eq) and fe / Y-butyl 4-aminoisoindoline-2-carboxylate (CAS 871013- 98-8, 259 mg, 1.11 mmol, 1.0 eq) were dissolved in DCM (6.0 mL) and / V, / V-diisopropylethylamine (0.39 mL, 2.21 mmol, 2.0 eq) was added. The reaction mixture was stirred at room temperature for 2 h. The resulting mixture was diluted with DCM (30 mL) and washed with water (30 mL). The aqueous phase was extracted with DCM (2 x 30 mL) and the combined organic extracts were washed with brine (30 mL), filtered through a hydrophobic frit, and concentrated under reduced pressure. The crude residue was purified by chromatography on silica, eluting with a gradient of 0 - 100% ethyl acetate in cyclohexane to afford the title compound (298 mg, Yield: 59%) which was used without further purification.Step 5: (E)- / V-isoindolin-4-yl-4-(isopropylamino)but-2-enamide bis trifluoroacetate

[0335] To a solution of te / Y-butyl 4-[[(E)-4-[fe / Y-butoxycarbonyl(isopropyl)amino]but-2- enoyl]amino]isoindoline-2-carboxylate (Step 4) (298 mg, 0.648 mmol, 1 .0 eq) in DCM (3.0 mL)was added trifluoroacetic acid (1 .0 mL, 13.2 mmol, 20.4 eq). The reaction mixture was stirred at room temperature for 2 h then concentrated under reduced pressure to afford the title compound (316 mg, Yield: 100%) which was used without further purification.Step 6: (E)- / V-(2-(3-Chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-4-(isopropylamino)but-

[0336] To a suspension of 3-chloro-6-hydroxy-4-methoxy-2-methylbenzoic acid (Intermediate D) (140 mg, 0.648 mmol, 1.0 eq), (E)- / V-isoindolin-4-yl-4-(isopropylamino)but-2-enamide bis trifluoroacetate (Step5) (316 mg, 0.648 mmol), 1 -hydroxybenzotriazole hydrate (99 mg, 0.648 mmol, 1.0 eq) and / V-(3- dimethylaminopropyl)- / '-ethylcarbodiimide hydrochloride (249 mg, 1.30 mmol, 2.0 eq) in DCM (7.0 mL) at room temperature was added 4-methylmorpholine (0.71 mL, 6.48 mmol, 10.0 eq). The reaction mixture was stirred for 3 days under an inert atmosphere. The resulting mixture was diluted with DCM (10 mL) and washed with water (10 mL), brine (10 mL), filtered through a hydrophobic frit, and the solvent concentrated under reduced pressure. The crude residue was purified by reverse phase preparative HPLC (Method: Prep-LC-1) then preparative SFC (Method: SFC-2) to afford the title compound (8.3 mg, Yield: 2.8%).

[0337] 1H NMR (400 MHz, DMSO) 6 1.02 - 0.95 (m, 6H), 2.18 (s, 3H), 2.78 - 2.65 (m, 1 H), 3.28 (d, J=11 .7 Hz, 2H), 3.82 - 3.80 (m, 3H), 4.43 (dd, J=4.3, 14.9 Hz, 1 H), 4.55 (dd, J=7.1 , 14.8 Hz, 1 H), 4.82 - 4.75 (m, 2H), 6.45 - 6.23 (m, 1 H), 6.53 (s, 1 H), 6.90 - 6.75 (m, 1 H), 7.17 - 7.00 (m, 1 H), 7.31 - 7.22 (m, 1 H), 7.67 (dd, J=8.0, 13.6 Hz, 1 H), 10.45 - 9.35 (m, 2H).

[0338] LCMS (LC-Method 1): 2.95 min, MS:ES+ 458.45 (M+1).

[0339] The compounds in the following table were prepared analogously to Example 14, Step 6 from the indicated intermediates.Example 15: (E)- / V-(5-Chloro-2-(3-chloro-4-hvdroxy-2-methylbenzoyl)isoindolin-4-yl)-4-(dimethylamino)- / V-methylbut-2-enamideStep 1 : Methyl 3-chloro-4-hydroxy-2-methylbenzoate

[0340] Performed in 2 parallel batches, each of 0.5 g scale: A stirred solution 4-bromo-2-chloro-3- methylphenol (CAS: 1799612-08-0, 0.5 g, 2.26 mmol, 1 eq) in MeOH: DMF (10 mL) at room temperature was degassed with N2 gas for 10 mins and treated with TEA (0.683 g, 6.76 mol, 3 eq) and PdCl2(dppf) (0.33 g, 0.45 mmol, 0.2 eq). The reaction mixture was placed under a positive pressure of CO(g) (20 kg / cm2) and heated to 120°C for 16 h. After cooling to room temperature, the resulting mixture was poured into ice cold water (50 mL) and extracted with EtOAc (3 x 100 mL). The combined organic extracts were dried over Na2SC>4, filtered, and concentrated under reduced pressure. The crude material was purified by chromatography on silica (product eluted at 15% EtOAc in hexane) yielding methyl 3-chloro-4-hydroxy-2- methylbenzoate as a white solid (0.55 g, Yield: 61 %).

[0341] 1H NMR (DMSO, 400 MHz): 6 2.56 (s, 3H), 3.78 (s, 3H), 6.90 (d, J= 8.4 Hz, 1 H), 7.66 (d, J= 8.4 Hz, 1 H), 10.94 (s, br, 1 H).

[0342] LCMS (Method N): 1.936 min, MS: ES+ 199.1 , 201 .1 (M, M+2).Step 2: 3-Chloro-4-hydroxy-2-methylbenzoic acid

[0343] To a stirred solution of methyl 3-chloro-4-hydroxy-2-methylbenzoate (Step 1) (0.5 g, 2.5 mmol, 1 eq) in MeOH: H2O (1 :1) (5 mL) was added KOH (2.8 g, 50 mmol, 20 eq) and the reaction mixture heated to 60°C for 30 mins. The resulting mixture was poured into ice cold water (50 mL), neutralized with dilute HCI (2 mL) and extracted with EtOAc (3 x 50 mL). The combined organic extracts were dried over Na2SO4, filtered and concentrated under reduced pressure to yield 3-chloro-4-hydroxy-2-methylbenzoic acid as an off-white solid (0.5 g, Yield: 98%). The crude material was used in the next step without further purification.

[0344] 1H NMR (DMSO, 400 MHz): 6 2.57 (s, 3H), 6.87 (d, J= 8.8 Hz, 1 H), 7.67 (d, J= 8.8 Hz, 1 H), 10.82 (s, 1 H), 12.49 (s, br, 1 H).

[0345] LCMS (Method N): 1.659 min, MS: ES+ 185.0, 187.0 (M, M+2).Step 3: te / Y-Butyl 5-chloro-4-(methylamino)isoindoline-2-carboxylate

[0346] To a stirred solution of fe / Y-butyl-4-(methylamino)isoindoline-2-carboxylate (Intermediate H) (5.0 g, 20.16 mmol, 1.0 eq.) in MeCN (50 mL) at room temperature was added / V-chlorosuccinimide (2.68 g, 20.16 mmol, 1 .0 eq.); the reaction mixture was stirred for 16 h. The resulting mixture was filtered and the filtrate concentrated under reduced pressure. The crude material was purified by chromatography on silica (product eluted at 5.0% EtOAc in hexane) to give the title compound (1.3 g, Yield: 22.9%). te / Y-Butyl 4- chloro-7-(methylamino)isoindoline-2-carboxylate was also isolated as a by-product (eluted at 10.0% EtOAc in hexane, 3.2 g, Yield: 56.3%).

[0347] 1H NMR (DMSO-d6, 400 MHz): 6 1.45 (s, 9H), 2.96 - 2.92 (m, 3H). 4.45 (d, J= 9.6 Hz, 2H), 4.81 (s, 2H), 6.56 (t, J= 17.6 Hz, 1 H), 7.16 (d, J= 7.6 Hz, 1 H).

[0348] LCMS (Method A): 2.273 min, MS: ES+ 226.8 (M-56).Step 4: te / Y-Butyl (E)-5-chloro-4-(4-(dimethylamino)- / V-methylbut-2-enamido)isoindoline-2-carboxylateMW, 15 mins

[0349] The title compound was prepared from (E)-4-(dimethylamino) but-2-enoic acid (CAS: 848133- 35-7) and fe / Y-butyl 5-chloro-4-(methylamino)isoindoline-2-carboxylate (Step 3) analogously to Example 9.

[0350] 1H NMR (DMSO-d6, 400 MHz): 5 1.45 (s, 9H), 2.24 (s, 6H), 3.12 (s, 3H), 3.17 - 3.19 (m, 2H), 4.36 - 4.42 (t, J=11 .2 Hz, J= 26 Hz, 1 H), 4.62 - 4.66 (m, 3H), 5.77 - 5.83 (m, 1 H), 6.67 - 6.74 (m, 1 H), 7.43- 7.46 (m, 1 H), 7.59 - 7.61 (d, J= 8 Hz, 1 H).

[0351] LCMS (Method N): 1.849 min, MS ES+: 394 (M+1).Step 5: (E)- / V-(5-Chloroisoindolin-4-yl)-4-(dimethylamino)- / V-methylbut-2-enamide hydrochloride

[0352] To a cooled (0°C) solution of te / Y-butyl (E)-5-chloro-4-(4-(dimethylamino)- / V-methylbut-2- enamido)isoindoline-2-carboxylate (Step 4) (0.4 g, 1.01 mmol, 1 eq) in DCM (4 mL) was added dropwise 4M HCI in dioxane (4 mL) and the reaction mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure and the crude material was triturated with diethyl ether (2 x 20 mL) followed by high vacuum drying to yield (E)- / V-(5-chloroisoindolin-4-yl)-4-(dimethylamino)- / V- methylbut-2-enamide hydrochloride (0.3 g, Yield: 89%).

[0353] 1H NMR (DMSO-d6, 400 MHz): 6 2.63 (s, br, 6H), 3.14 (s, 3H), 3.74 - 3.79 (m, 2H), 4.39 - 4.42 (d, J= 14 Hz, 1 H), 4.57 (s, br, 3H), 6.05 (d, J= 15.2 Hz, 1 H), 6.71 - 6.78 (m, 1 H), 7.51 (d, J= 8 Hz, 1 H), 7.67 (d, J= 8.0 Hz, 1 H), 10.06 (s, br, 1 H), 10.44 (s, br, 1 H), 10.65 (s, br, 1 H).

[0354] LCMS (Method O): 1.449 min, MS ES+: 294.12 (M+1).Step 6: (E)- / V-(5-Chloro-2-(3-chloro-4-hydroxy-2-methylbenzoyl)isoindolin-4-yl)-4-(dimethylamino)- / V- methylbut-2-enamide. , , NMM, DMF, rt, 15 minutes

[0355] The title compound was prepared from 3-chloro-4-hydroxy-2-methylbenzoic acid (Step 2) and (E)- / V-(5-chloroisoindolin-4-yl)-4-(dimethylamino)- / V-methylbut-2-enamide hydrochloride (Step 5) analogously to Example 4.

[0356] 1H NMR (DMSO-d6, 400 MHz): 6 2.01 (d, J= 2.0 Hz, 6H), 2.20 - 2.24 (m, 3H), 2.85 - 2.89 (m, 2H), 3.01 and 3.15 (2 singlets, 3H), 4.54 - 4.91 (m, 4H), 5.68 (dd, J= 24.4 Hz, J= 24.4 Hz, 1 H), 6.60 - 6.74 (m, 1 H), 6.89 (dd, J= 14.4 Hz, J= 14.8 Hz, 1 H), 7.08 - 7.13 (m, 1 H), 7.33 - 7.52 (m, 1 H), 7.60 (q, J= 14.0 Hz, J= 14.4 Hz, 1 H), 10.42 (bs, 1 H).

[0357] LCMS (Method O): 5.778 min, MS ES+: 462.1 , 464.1 (M, M+2).

[0358] HPLC (Method H): 3.563 min.

[0359] Prep. HPLC purification and method of analysis: Chromatographic separation and isolation were conducted on a Waters 2545 binary pump with Waters 2489 UV detector. Column Waters X-Bridge C18 (250 mm x 19 mm x 5pm); compounds eluted with: Mobile Phase A: 0.05% NH3 + 5mm ammonium bicarbonate in water; Mobile Phase B: Acetonitrile with a gradient of T = 0.01 min (72% A, 28% B); gradient to T = 24.00 min (72% A, 28% B); T = 24.01 min (00% A, 100% B), gradient to T = 27.00min (00% A, 100% B); T = 27.01 min (72% A, 28% B); gradient to T = 35.00 min (72% A, 28% B); Flow rate= 09 ml / min; analysis time 35.00 min.Example _ 16: _ (E)- / V-(2-(3-Chloro-6-hvdroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-3-(1- methylpyrrolidin-2-yl)acrylamideStep 1 : te / Y-Butvl (E)-4-(3-(1-methylpyrrolidin-2-yl) acrylamido) isoindoline-2-carboxylateDCM, O°C-rt, 3h

[0360] The title compound was prepared from (E)-3-(1-methylpyrrolidin-2-yl)acrylic acid (Example 11 , Step 2) and fe / Y-butyl 4-aminoisoindoline-2-carboxylate (CAS: 871013-98-8) analogously to Example 6, Step 1 .

[0361] 1H NMR (DMSO-d6, 400 MHz): 6 1.46 (s, 9H), 1.62 (s, br, 1 H), 1.72 - 1.77 (m, 2H), 2.04 - 2.06 (m, 1 H), 2.28 - 2.33 (m, 3H), 2.91 - 3.11 (m, 2H), 4.47 - 4.61 (m, 4H), 6.45 - 6.47 (m, 1 H), 6.65 (s, br, 1 H), 7.08 - 7.10 (m, 1 H), 7.26 (t, J= 7.6 Hz, 1 H), 7.65 - 7.67 (m, 1 H), 9.68 (bs, 1 H).

[0362] LCMS (Method N): 1.299 min, MS: ES+ 372.2 (M+1).Step 2: (£)- / V-(lsoi ndol i n-4-y l)-3-(1 -methylpyrrolidin-2-yl)acrylamide hydrochloride

[0363] The title compound was prepared from te / Y-butyl (E)-4-(3-(1-methylpyrrolidin-2-yl)acrylamido) isoindoline-2-carboxylate (Step 1) and 4M HCI analogously to Example 6, Step 2.

[0364] 1H NMR (DMSO-d6, 400 MHz, D2O exchange): 6 1.91 - 2.07 (m, 3H), 2.26 (m, 1 H), 2.75 (s, 3H), 3.10 - 3.16 (m, 2H), 4.08 (s, br 1 H), 4.48 - 4.52 (m, 4H), 6.65 (d, J= 15.2 Hz, 1 H), 6.80 - 6.92 (m, 1 H), 7.17 - 7.23 (m, 1 H), 7.37 (t, J= 7.6 Hz, 1 H), 7.55 (d, J= 8.0 Hz,1 H).

[0365] LCMS (Method O): 1.083 min, MS: ES- 270.3, (M-1).Step 3: (E)- / V-(2-(3-Chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-3-(1-methylpyrrolidin-2- yl)acrylamideNMM, DMF, rt, 1 h

[0366] The title compound was prepared from 3-chloro-6-hydroxy-4-methoxy-2-methylbenzoic acid (Intermediate D) and (E)- / V-(isoindolin-4-yl)-3-(1-methylpyrrolidin-2-yl)acrylamide hydrochloride (Step 2) analogously to Example 4.

[0367] HT1H NMR (DMSO-d6, 400 MHz, 348K): 6 1.15 - 1.62 (m, 1 H), 1.71 - 1.77 (m, 2H), 1.96 - 2.07 (m, 1 H), 2.20 - 2.26 (m, 4H), 2.26 - 2.29 (m, 3H), 2.75 - 2.84 (m, 1 H), 2.99 - 3.08 (m, 1 H), 3.83 (s, 3H), 4.44 (t, J= 13.2 Hz, 1 H), 4.59 (d, J= 14.8 Hz, 1 H), 4.82 (d, J= 12.0 Hz, 2H), 6.29 - 6.42 (m, 1 H), 6.58 (m, 1 H), 6.61 - 6.73 (m, 1 H), 7.02 - 7.16 (m, 1 H), 7.23 - 7.30 (m, 1 H), 7.63 - 7.73 (m, 1 H), 8.19 (bs, 1 H), 9.30 - 9.52 (m, 1 H).

[0368] LCMS (Method N): 1.207 min, MS: ES+ 470.1 , 472.1 (M, M+2).

[0369] HPLC (Method I): 6.063 min.

[0370] Prep. HPLC purification and method of analysis: Chromatographic separation and isolation were conducted with Waters 2545 Quaternary system with waters 2489 UV Detector. Column Sunfire Prep C18 (250 x 19 mm, 5um); compounds eluted with, Mobile Phase A: 0.05% Formic acid in water, Mobile Phase B: Acetonitrile: H2O: THF (8:2:1) with a gradient of T = 0.01 (85% A, 15% B); gradient to T = 25.00 min (76% A, 24% B); T =35.00 min (76% A, 24% B) gradient to T = 35.01 min (0% A, 100% B); T = 37.00 min (0% A, 100% B); gradient to T = 37.01 min (85% A, 15% B); T = 40.00 min (85% A, 15% B); Flow rate= 10ml / min; analysis time 40.00 min.Example 16.1 : (E)- / V-(2-(3-Chloro-2-fluoro-6-hvdroxy-4-methoxybenzoyl)isoindolin-4-yl)-3-(1- methylpyrrolidin-2-yl)acrylamideNMM, DMF, rt, 1 h

[0371] A stirred solution of 3-chloro-2-fluoro-6-hydroxy-4-methoxybenzoic acid (0.15 g, 0.68 mmol, 1.0 eq. Intermediate U) in DMF (1.5 mL) at room temperature was treated with EDC.HCI (0.196 g, 1.02 mmol, 1.5 eq) and HOAt (0.092 g, 0.68 mmol, 1 eq) and stirred for 10mins. (E)- / V-(isoindolin-4-yl)-3-(1 - methylpyrrolidin-2-yl)acrylamide hydrochloride (0.313 g, 1 .02 mmol, 1 .5 eq, Example 16 Step 2) and NMM (0.375 mL, 3.40 mmol, 5.0 eq) were added, and the resulting mixture stirred at room temperature for 16h. The reaction mixture was poured into ice cold water (100 mL), extracted using EtOAc (3 x 80 mL) and the combined organic layer dried over Na2SO4, filtered, and concentrated under reduced pressure to givecrude material (0.419 g) which was purified by reverse phase column chromatography (product eluted in 25% water : acetonitrile) followed by Prep. HPLC (see conditions below) yielding the title compound as a white solid (0.043 g, Yield: 13%, 0.090 mmol).

[0372] 1H NMR (DMSO-d6, 400 MHz, 348K): 6 ppm 1 .60 (bs, 1 H), 1 .76 (s, br, 2H), 2.05 (s, br, 1 H), 2.14 and 2.23 (2 singlets, 3H), 2.22 - 2.28 (m, 2H), 2.81 - 2.89 (m, 1 H), 3.88 (s, 3H), 4.63 (s, 2H), 4.82 (d, J = 12.4 Hz, 2H), 6.26 - 6.42 (m, 1 H), 6.56 (s, 1 H), 7.04 - 7.17 (m, 1 H), 7.25 - 7.27 (m, 1 H), 7.64 - 7.72 (m, 1 H), 8.29 - 8.78 (m, 1 H), 9.32 and 9.54 (2 singlets, 1 H).

[0373] LCMS (Method N): 1.697 min, MS: ES+ 474.3 (M+1).

[0374] HPLC (Method H): 4.19 min, 254nm.

[0375] Prep. HPLC purification: Chromatographic separation and isolation were conducted using a Shimadzu LC20SP with a UV detector; column: X-Bridge Prep C18 (250 x 19 mm x 5um), compounds eluted with Mobile Phase A: 5mM ammonium bicarbonate + 0.05% NH3 in Merck water: Mobile Phase B: Acetonitrile: MeOH (50:50) with a gradient of T = 0.01 min (70% A, 30% B); gradient to T = 16.00 min (55% A, 45% B); T = 22.00 min (50% A, 50% B); gradient to T = 22.01 min (0% A, 100% B); T = 24.00 min (0% A, 100% B); T = 24.01 min (70% A, 30% B); T = 26.00 min (70% A, 30% B); Flow rate= 15 ml / min; analysis time 26 min.Example 17: (E)- / V-(2-(3-Chloro-6-hvdroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-3-(pyrrolidin-2- vDacrylamideStep 1 : Ethyl (E)-3-(pyrrolidin-2-yl) acrylate hydrochloride

[0376] A cooled (0°C) solution of te / Y-butyl (E)-2-(3-ethoxy-3-oxoprop-1-en-1-yl)pyrrolidine-1- carboxylate (Intermediate M) (8 g, 29.73 mmol, 1 eq.) in DCM (80 mL) was treated with 4M HCI in dioxane (80 mL) and the resulting reaction mixture was stirred at room temperature for 1 h. The mixture was concentrated under reduced pressure yielding ethyl (E)-3-(pyrrolidin-2-yl) acrylate hydrochloride as a brown oil (8 g, Yield: quantitative). The crude material was used in the next step without further purification.

[0377] 1H NMR (DMSO, 400 MHz): 6 1 .23 (t, J= 7.2 Hz, 3H), 1 .73 - 1 .78 (m, 1 H), 1 .89 - 2.00 (m, 2H), 2.13 - 2.18 (m, 1 H), 3.17 - 3.25 (m, 2H), 4.1 1 - 4.23 (m, 3H), 6.19 - 6.23 (m, 1 H), 6.93 - 6.98 (m, 1 H), 9.22 (s, 1 H), 9.83 (s, 1 H).

[0378] LCMS (Method N): 0.872 min, MS: ES+ 170.1 (M+1).Step 2: Benzyl (E)-2-(3-ethoxy-3-oxoprop-1-en-1-yl) pyrrolidine-1 -carboxylate

[0379] A cooled (0°C) solution of ethyl (E)-3-(pyrrolidin-2-yl) acrylate hydrochloride (Step 1) (8 g, 39.02 mmol, 1 eq.) in THF (80 mL) was treated with TEA (7.84 g, 77.66 mmol, 2.0 eq.) and the mixture stirred for 5 mins. Benzylchloroformate (9.9 g, 58.25 mmol, 1 .5 eq) was added dropwise at 0°C and the reaction mixture was allowed to warm to room temperature stirring for 1 h. The resulting mixture was poured into ice cold water (100 mL) and extracted with EtOAc (3 x 100 mL). The combined organic extracts were dried over Na2SC>4, filtered, and concentrated under reduced pressure. The crude material was purified twice by chromatography on silica (product eluted at 20% EtOAc: Hexane) yielding benzyl-(E)-2-(3-ethoxy-3- oxoprop-1-en-1-yl) pyrrolidine-1 -carboxylate as a yellow oil (8 g, Yield: 80%).

[0380] 1H NMR (DMSO, 400 MHz): 6 1.21 (t, J= 6.8 Hz, 3H), 1.76 - 1 .82 (m, 3H), 2.05 - 2.07 (m, 1 H), 3.38 - 3.46 (m, 2H), 4.11 (q, J= 7.2 Hz, 2H), 4.45 - 4.50 (m, 1 H), 5.08 - 5.18 (m, 2H), 5.76 (t, J= 14.8 Hz, 1 H), 6.81 (dd, J= 5.6 Hz, 15.6 Hz, 1 H), 7.29 - 7.37 (m, 5H).

[0381] LCMS (Method N): 1.78 min, MS: ES+ 304.3 (M+1).Step 3: (E)-3-(1-((Benzyloxy)carbonyl) pyrrolidin-2-yl) acrylic acid

[0382] To a stirred solution of benzyl-(E)-2-(3-ethoxy-3-oxoprop-1-en-1-yl) pyrrolidine-1 -carboxylate (Step 2) (8 g, 26.40 mmol, 1 eq.) in IPA: water (1 :1) (80 mL) was added NaOH (10.56 g, 264.0 mmol, 10 eq.) and the reaction mixture was stirred at room temperature for 16 h. The resulting mixture was poured into ice cold water (100 mL), neutralized with sat. KHSC solution and extracted with EtOAc (3 x 100 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated under reduced pressure yielding (E)-3-(1-((benzyloxy)carbonyl) pyrrolidin-2-yl) acrylic acid as a yellow oil (8 g, Yield: Quantitative). The crude was used in the next step without further purification.

[0383] 1H NMR (DMSO, 400 MHz): 6 1.75 - 1 .82 (m, 3H), 2.04 - 2.07 (m, 1 H), 3.37 - 3.43 (m, 2H), 4.44 - 4.49 (m, 1 H), 4.99 - 5.11 (m, 2H), 5.70 (d, J= 12.4 Hz, 1 H), 6.72 (d, J= 15.2 Hz, 1 H), 7.23 - 7.37 (m, 5H), 12.35 (bs, 1 H).

[0384] LCMS (Method N): 1.423 min, MS: ES+ 276.1 (M+1).Step 4: te / Y-Butvl (E)-4-(3-(1-((benzyloxy)carbonyl) pyrrolidin-2-yl) acrylamido) isoindoline-2-carboxylate

[0385] The title compound was prepared from (E)-3-(1-((benzyloxy)carbonyl) pyrrolidin-2-yl) acrylic acid (Step 3) and fe / Y-butyl-4-aminoisoindoline-2-carboxylate (CAS: 871013-98-8) analogously to Example 6, Step 1 .

[0386] 1H NMR (DMSO, 400 MHz): 6 1 .46 (s, 9H), 1 .79 - 1 .86 (m, 3H), 2.08 - 2.10 (m, 1 H), 3.43 - 3.44 (m, 2H), 4.51 - 4.62 (m, 5H), 5.00 - 5.16 (m, 2H), 6.27 (d, J= 15.2 Hz, 1 H), 6.73 - 6.76 (m, 1 H), 7.10 (t, J= 6.4 Hz, 1 H), 7.25 - 7.38 (m, 6H), 7.65 - 7.67 (m, 1 H), 9.62 (s, 1 H).

[0387] LCMS (Method N): 1.970 min, ES+ 509.3 (M+18).Step 5: Benzyl-(E)-2-(3-(isoindolin-4-ylamino)-3-oxoprop-1 -en-1 -yl) pyrrolidine-1 -carboxylate hydrochloride

[0388] The title compound was prepared from fe / Y-butyl (E)-4-(3-(1-((benzyloxy)carbonyl) pyrrolidin-2- yl) acrylamido) isoindoline-2-carboxylate (Step 4) and 4M HCI in dioxane analogously to Example 6, Step 2.

[0389] 1H NMR (DMSO, 400 MHz): 6 1.78 - 1 .86 (m, 3H), 2.08 - 2.10 (m, 1 H), 3.42 - 3.53 (m, 2H), 3.57 (m, 1 H), 4.44 - 4.54 (m, 4H), 5.00 - 5.18 (m, 2H), 6.18 (d, J= 14 Hz, 1 H), 6.75 - 6.78 (m, 1 H), 7.21 (d, J= 7.6 Hz, 1 H), 7.32 - 7.37 (m, 6H), 7.43 - 7.48 (m, 1 H), 9.5 (s, 1 H), 10.02 (d, J= 12.8 Hz, 1 H).

[0390] LCMS (Method O): 1.332 min, MS: ES+ 391.9 (M+1).Step 6: Benzyl-(E)-2-(3-((2-(3-chloro-6-hydroxy-4-methoxy-2-methylbenzoyl) isoindolin-4-yl) amino)-3- oxoprop-1 -en-1 -yl) pyrrolidine-1 -carboxylateNMM, DMF, rt, 16h

[0391] The title compound was prepared from 3-chloro-6-hydroxy-4-methoxy-2-methylbenzoic acid (Intermediate D) and benzyl (E)-2-(3-(isoindolin-4-ylamino)-3-oxoprop-1-en-1-yl) pyrrolidine-1 -carboxylate hydrochloride (Step 5) analogously to Example 4.

[0392] 1H NMR (DMSO, 400 MHz): 6 1.81 - 1 .87 (m, 3H), 2.05 - 2.13 (m, 1 H), 2.18 (s, 3H), 3.36 - 3.43 (m, 2H), 3.77 - 3.82 (m, 3H), 4.41 - 4.58 (m, 2H), 4.77 - 4.82 (m, 2H), 4.94 - 5.19 (m, 2H), 6.22 (dd, J= 15.2 Hz, 51 .2 Hz, 1 H), 6.55 (s, 1 H), 6.67 - 6.79 (m, 1 H), 7.10 (dd, J= 7.6 Hz, 49.2 Hz, 1 H), 7.25 - 7.39 (m, 5H), 7.66 - 7.74 (m, 1 H), 9.51 - 9.71 (m, 1 H), 10.04 (s, 1 H).

[0393] LCMS (Method N): 1.704 min, ES+ 590.2 (M).Step 7: (E)- / V-(2-(3-Chloro-6-hydroxy-4-methoxy-2-methylbenzoyl)isoindolin-4-yl)-3-(pyrrolidin-2- yl)acrylamide

[0394] The title compound was prepared from benzyl (E)-2-(3-((2-(3-chloro-6-hydroxy-4-methoxy-2- methylbenzoyl) isoindolin-4-yl) amino)-3-oxoprop-1-en-1-yl) pyrrolidine-1 -carboxylate (Step 6) and anhydrous AlCh analogously to Example 6, Step 4.

[0395] HT1H NMR (DMSO, 400 MHz, 348K): 6 1.40 - 1 .50 (m, 1 H), 1 .67 - 1 .73 (m, 2H), 1 .89 - 2.01 (m, 1 H), 2.21 (s, 3H), 2.81 - 2.95 (m, 2H), 3.65 - 3.73 (m, 1 H), 3.83 (s, 3H), 4.44 (t, J= 12.8 Hz, 1 H), 4.57 - 4.62 (m, 1 H), 4.81 (d, J= 15.2 Hz, 2H), 6.32 (dd, J= 15.2 Hz, 51.6 Hz, 1 H), 6.56 (s, 1 H), 6.70 - 6.84 (m, 1 H), 7.08 (dd, J= 7.6 Hz, 52.4 Hz, 1 H), 7.23 - 7.30 (m, 1 H), 7.62 - 7.68 (m, 1 H) 9.21 - 9.45 (m, 1 H).

[0396] LCMS (Method N): 1.205 min, ES+ 456.1 (M).

[0397] HPLC (Method H): 4.144 min.Example 18: (E)- / V-(2-(5-Chloro-2-hvdroxy-4-methoxybenzoyl)isoindolin-4-yl)-3-(pyrrolidin-2-yl)acrylamideStep 1 : Benzyl-(E)-2-(3-((2-(5-chloro-2-hydroxy-4-methoxybenzoyl) iso i ndol i n-4-y I) amino)-3-oxoprop-1 - en-1 -yl) pyrrolidine-1 -carboxylateNMM, DMF, rt, 16h

[0398] The title compound was prepared from 5-chloro-2-hydroxy-4-methoxybenzoic acid (CAS: 1378866-39-7) and benzyl(E)-2-(3-(isoindolin-4-ylamino)-3-oxoprop-1 -en-1 -yl) pyrrolidine-1 -carboxylate hydrochloride (Example 17, Step 5) analogously to Example 4.

[0399] 1H NMR (DMSO, 400 MHz): 6 1.82 - 1 .85 (m, 3H), 2.09 (bs, 1 H), 3.36 - 3.50 (m, 3H), 3.81 - 3.85 (m, 1 H), 4.51 - 4.57 (m, 1 H), 4.70 - 4.81 (m, 3H), 5.01 - 5.13 (m, 2H), 6.15 - 6.31 (m, 1 H), 6.55 (s, 1 H), 6.67 - 6.78 (m, 1 H), 7.05 - 7.17 (m, 1 H), 7.27 - 7.39 (m, 7H), 7.67 - 7.68 (m, 1 H), 9.57 - 9.71 (m, 1 H), 10.46 - 10.60 (m, 1 H).

[0400] LCMS (Method N): 1.815 min, ES+ 576.1 (M).Step 2: (E)- / V-(2-(5-Chloro-2-hydroxy-4-methoxybenzoyl)isoindolin-4-yl)-3-(pyrrolidin-2-yl)acrylamide

[0401] The title compound was prepared from benzyl (E)-2-(3-((2-(5-chloro-2-hydroxy-4- methoxybenzoyl) isoindolin-4-yl) amino)-3-oxoprop-1-en-1-yl) pyrrolidine-1 -carboxylate (Step 1) and anhydrous AlCh analogously to Example 6, Step 4.

[0402] HT1H NMR (DMSO, 400 MHz, 348K): 6 1.43 - 1 .49 (m, 1 H), 1 .68 - 1 .75 (m, 2H), 1 .90 - 1 .99 (m, 1 H), 2.82 - 2.93 (m, 2H), 3.68 - 3.71 (m, 1 H), 3.86 (s, 3H), 4.80 (d, J= 10.4 Hz, 4H), 6.33 (d, J= 15.2 Hz, 1 H), 6.69 (s, 1 H), 6.78 (dd, J= 5.6 Hz, 14.8 Hz, 1 H), 7.09 - 7.11 (m, 1 H), 7.27 (t, J= 7.6 Hz, 1 H), 7.36 (s, 1 H), 7.63 (d, J= 8.0 Hz, 1 H), 9.43 (s, 1 H).

[0403] LCMS (Method N): 1.216 min, ES+ 442.1 (M).

[0404] HPLC (Method H): 4.153 min.Example 19: (E)- / V-(5-Chloro-2-(5-chloro-2-hvdroxy-4-methoxybenzoyl)isoindolin-4-yl)-3-(pyrrolidin-2- vDacrylamideStep 1 : fe / Y-Butyl(E)-4-(3-(1-((benzyloxy)carbonyl)pyrrolidin-2-yl)acrylamido)-5-chloroisoindoline-2- carboxylate

[0405] The title compound was prepared from (E)-3-(1-((benzyloxy)carbonyl) pyrrolidin-2-yl)acrylic acid (Example 17, Step 3) and fe / Y-butyl-4-amino-5-chloroisoindoline-2-carboxylate (Example 3, Step 1) analogously to Example 6, Step 1 .

[0406] 1H NMR (DMSO, 400 MHz): 6 1 .39 (s, br, 1 H), 1 .45 (s, 9H), 1 .79 - 1 .86 (m, 2H), 2.07 (s, br, 2H), 3.36 - 3.44 (m, 1 H), 4.42 - 4.51 (m, 3H), 4.61 (s, br, 2H), 5.03 - 5.14 (m, 2H), 6.23 (d, J= 14.8 Hz, 1 H), 6.74 - 6.75 (m, 1 H), 7.25 - 7.38 (m, 5H), 7.45 (t, J= 8.4 Hz, 2H), 9.76 (d, J= 2.4 Hz, 1 H), 9.74 - 9.94 (m, 1 H).

[0407] LCMS (Method N): 1.941 min, ES+ 543.2 (M+18).Step 2: Benzyl-(E)-2-(3-((5-chloroisoindolin-4-yl)amino)-3-oxoprop-1-en-1-yl)pyrrolidine-1 -carboxylate hydrochloride

[0408] The title compound was prepared from te / Y-butyl (E)-4-(3-(1-((benzyloxy)carbonyl)pyrrolidin-2- yl)acrylamido)-5-chloroisoindoline-2-carboxylate (Step 1) and 4M HCI in dioxane analogously to Example 6, Step 2 and used directly in the next step.Step 3: Benzyl (E)-2-(3-((5-chloro-2-(5-chloro-2-hydroxy-4-methoxybenzoyl)isoindolin-4-yl)amino)-3- oxoprop-1 -en-1 -y I) py rro lid i ne- 1 -carboxylateNMM, DMF, rt, 16h

[0409] The title compound was prepared from 5-chloro-2-hydroxy-4-methoxybenzoic acid (CAS: 1378866-39-7) and benzyl(E)-2-(3-((5-chloroisoindolin-4-yl)amino)-3-oxoprop-1 -en-1 -y I) py rro I id ine- 1 - carboxylate hydrochloride (Step 2) analogously to Example 4.

[0410] 1H NMR (DMSO, 400 MHz): 6 1 .66 - 1 .83 (m, 3H), 2.07 - 2.09 (m, 1 H), 3.43 - 3.50 (m, 2H), 3.75 - 3.84 (m, 3H), 4.51 - 4.52 (m, 1 H), 4.60 - 4.65 (m, 2H), 4.75 - 4.87 (m, 2H), 4.97 - 5.19 (m, 2H), 6.11 - 6.28 (m, 1 H), 6.64 - 6.66 (m, 1 H), 6.76 - 6.79 (m, 1 H), 7.22 - 7.38 (m, 7H), 7.46 - 7.50 (m, 1 H), 9.86 - 10.01 (m, 1 H), 10.46 - 10.55 (m, 2H).

[0411] LCMS (Method N): 2.29 min, ES+ 610.1 (M).Step 4: (E)- / V-(5-Chloro-2-(5-chloro-2-hydroxy-4-methoxybenzoyl)isoindolin-4-yl)-3-(pyrrolidin-2- yl)acrylamide

[0412] The title compound was prepared from benzyl (E)-2-(3-((5-chloro-2-(5-chloro-2-hydroxy-4- methoxybenzoyl)isoindolin-4-yl)amino)-3-oxoprop-1 -en-1 -y I) py rro lid i n e- 1 -carboxylate (Step 3) and anhydrous AlCh analogously to Example 6, Step 4.

[0413] 1H NMR (DMSO, 400 MHz): 6 1.38 - 1 .48 (m, 1 H), 1 .66 - 1 .75 (m, 2H), 1 .89 - 1 .93 (m, 1 H), 2.83 - 2.87 (m, 2H), 3.64 - 3.71 (m, 1 H), 3.83 (s, 3H), 4.57 - 4.63 (m, 2H), 4.74 - 4.82 (m, 2H), 6.20 - 6.37 (m, 1 H), 6.65 - 6.84 (m, 2H), 7.21 - 7.32 (m, 2H), 7.45 - 7.50 (m, 1 H), 9.79 - 9.94 (m, 1 H).

[0414] LCMS (Method N): 1.67 min, ES+ 476.1 , 478.1 (M, M+2).

[0415] HPLC (Method H): 4.15 min.Example 20: (E)- / V-(5-Chloro-2-(5-chloro-2-hvdroxy-4-methoxybenzoyl)isoindolin-4-yl)-3-(1- methylpyrrolidin-2-yl)acrylamide

[0416] To a stirred solution of (E)- / V-(5-chloro-2-(5-chloro-2-hydroxy-4-methoxybenzoyl)isoindolin-4-yl)- 3-(pyrrolidin-2-yl)acrylamide (Example 19) (0.23 g 0.48 mmol, 1 eq.) in DCE (2.3 mL) was added paraformaldehyde (0.07 g, 2.41 mmol, 5 eq) and the mixture was stirred at room temperature for 1.5 h. Sodium triacetoxyborohydride (0.30 g, 1 .44 mmol, 3 eq) was added portion wise and the reaction mixturewas stirred at room temperature for 16 h. The resulting mixture was poured into saturated NaHCCh solution (40 mL) and extracted with DCM (2 x 40 mL). The combined organic extracts were dried over Na2SC>4, filtered and concentrated under reduced pressure. The crude material was purified by reverse phase chromatography (product eluted at 58% acetonitrile in water) followed by preparative TLC (9:1 DCM: MeOH) and pure fraction distillation and lyophilization to yield (E)- / V-(5-chloro-2-(5-chloro-2-hydroxy-4- methoxybenzoyl)isoindolin-4-yl)-3-(1-methylpyrrolidin-2-yl)acrylamide as a white solid (0.018g, Yield: 8%).

[0417] 1H NMR (DMSO-d6, 400 MHz): 6 1.54 - 1.73 (m, 4H), 1.90 - 2.03 (m, 3H), 2.10 - 2.25 (m, 3H), 3.05 (s, br, 1 H), 3.82 (d, J= 6.00 Hz, 3H), 4.60 (d, J= 24.8 Hz, 2H), 4.76 - 4.82 (m, 2H), 6.20, 6.38 (dd, J= 15.2 Hz, J= 39.2 Hz, 1 H), 6.54 - 6.71 (m, 2H), 7.22 (d, J= 8.0 Hz, 1 H), 7.29 - 7.33 (m, 1 H), 7.44 - 7.49 (m, 1 H), 9.99 (s, 1 H), 10.54 - 10.57 (m, 1 H).

[0418] LCMS (Method N): 1.230 min, MS: ES+ 490.1 , 492.1 (M, M+2).

[0419] HPLC (Method H): 4.289 min.Example 21 : (E)- / V-(2-(3-Chloro-6-hvdroxy-2,4-dimethoxybenzoyl)isoindolin-4-yl)-3-(1 -methylpyrrolidin-2- vDacrylamideStep 1 : 3-Chloro-6-hydroxy-2,4-dimethoxybenzoic acid

[0420] To a cooled (0°C) solution of2-hydroxy-4,6-dimethoxybenzoic acid (CAS: 3187-19-7) (0.5 g, 2.52 mmol, 1 .0 eq) in THF (5 mL) were added portion wise p-toluene sulfonic acid (0.047 g, 0.25 mmol, 0.1 eq) and NCS (0.35 g, 2.64 mmol, 1 .05 eq). The reaction mixture was stirred at room temperature for 4 h. The resulting mixture was diluted with sat. solution of sodium thiosulfate (100 mL) and extracted using EtOAc (2 x 100 mL). The combined organic extracts were dried over Na2SC>4, filtered, and concentrated under reduced pressure. The crude material was purified by reverse phase column chromatography (product eluted in 50% ACN in water) yielding 3-chloro-6-hydroxy-2,4-dimethoxybenzoic acid as a white solid (0.4 g, Yield: 68%).

[0421] 1H NMR (DMSO-d6, 400 MHz): 6 3.75 (s, 3H), 3.85 (s, 3H), 6.48 (s, 1 H).

[0422] LCMS (Method N): 1.871 min, MS: ES+ 233.0 (M+1).Step 2: (E)- / V-(2-(3-Chloro-6-hydroxy-2,4-dimethoxybenzoyl)isoindolin-4-yl)-3-(1-methylpyrrolidin-2- yl)acrylamide

[0423] The title compound was prepared from 3-chloro-6-hydroxy-2,4-dimethoxybenzoic acid (Step 1) and (E)- / V-(isoindolin-4-yl)-3-(1-methylpyrrolidin-2-yl)acrylamide hydrochloride (Example 16, Step 2) analogously to Example 4.

[0424] 1H NMR (DMSO-d6, 400 MHz): 6 1.55 - 1.57 (m, 1 H), 1.68 - 1 .74 (m, 2H), 1.94 - 2.02 (m, 1 H), 2.13 - 2.21 (m, 3H), 2.67 - 2.77 (m, 1 H), 2.98 - 3.03 (m, 1 H), 3.75 (s, 3H), 3.81 (s, 3H), 4.51 - 4.58 (m, 2H), 4.76 - 4.79 (m, 2H), 6.28 - 6.40 (m, 1 H), 6.47 (s, 1 H), 6.56 - 6.69 (m, 1 H), 7.03 - 7.15 (m, 1 H), 7.24 - 7.31 (m, 1 H) 7.64 - 7.77 (m, 1 H), 9.46 - 9.71 (m, 1 H), 10.35 (bs, 1 H).

[0425] LCMS (Method N): 1.217 min, MS: ES+ 486.1 , 488.1 (M, M+2).

[0426] HPLC (Method I): 5.84 min.

[0427] Prep. HPLC purification and method of analysis: Chromatographic separation and isolation were conducted with Waters 2545 Quaternary system with Waters 2489 UV Detector. Column X Bridge C18 (250 x 19 mm ID, 5um); compounds eluted with: Mobile Phase A: 5mm ammonium bicarbonate + 0.05% NH3 in water; Mobile Phase B: Acetonitrile with a gradient of T = 0.01 (77% A, 23% B); gradient to T = 20.00 min (65% A, 35% B); T =22.00 min (65% A, 35% B) gradient to T = 22.01 min (0% A, 100% B); T = 24.00 min (0% A, 100% B); gradient to T = 24.01 min (77% A, 23% B); T = 30.00 min (77% A, 23% B); Flow rate= 12 mL / min; analysis time 30.00 min.Example 21 .1 (E)- / V-(2-(3-Chloro-6-hvdroxy-2,4-dimethoxybenzoyl)isoindolin-4-yl)-4-(dimethylamino)but-2-enamideNMM, DMF, rt, 1 h

[0428] A stirred solution of 3-chloro-6-hydroxy-2,4-dimethoxybenzoic acid (0.35 g, 1.5 mmol, 1.0 eq, Example 21 , Step 1) in DMF (3.5 mL) at room temperature was treated with EDC.HCI (0.43 g, 2.2 mmol, 1 .5 eq) and HOAt (0.20 g, 1 .5 mmol, 1 .0 eq) and stirred for 10 min. (E)-4-(dimethylamino)- / V-(isoindolin-4- yl)but-2-enamide dihydrochloride (Intermediate T) (0.634 g, 2.2 mmol, 1.5 eq) and NMM (1 mL, 9.0 mmol, 6.0 eq) were added and the resulting reaction mixture stirred at room temperature for 1 h. The reaction mixture was poured into ice cold water (125 mL) and extracted using EtOAc (2 x 100 mL). The combined organic layer was washed with 1 M HCI, dried over Na2SC>4, filtered, and concentrated under reducedpressure to give crude material (0.33 g) which was purified by prep. HPLC (see conditions below) yielding the title compound as a white solid (0.034 g, Yield: 5.2%).

[0429] 1H NMR (DMSO-d6, 400 MHz, 348K): 6 ppm 2.18 and 2.23 (2 singlets, 6H), 3.03 (d, J = 6 Hz, 1 H), 3.09 (d, J= 4.4 Hz, 1 H), 3.77 (s, 3H), 3.85 (s, 3H), 4.57 (s, 2H), 4.81 (d, J = 12 Hz, 2H), 6.27 - 6.43 (m, 1 H), 6.50 (s, 1 H), 6.67 - 6.82 (m, 1 H), 7.03 - 7.17 (m, 1 H), 7.24 - 7.31 (m, 1 H), 7.63 - 7.70 (m, 1 H), 9.32 and 9.52 (2 singlets, 1 H), 9.89 (bs, 1 H).

[0430] LCMS (Method N): 1.671 min, MS: ES+ 460.3, 462.3 (M, M+2).

[0431] HPLC (Method I): 5.85 min, 210 nm

[0432] Prep. HPLC purification: Chromatographic separation and isolation were conducted using a Waters 2545 Quaternary system with Waters 2489 UV Detector and X-Bridge Prep C18 (150 x 19 mm, 5um) column; compounds were eluted with: Mobile Phase A: 5mM ammonium bicarbonate + 0.05% NH3 in water, Mobile Phase B: Acetonitrile with a gradient of T = 0.00 (85% A, 15% B); gradient to T = 25.00 min (80% A, 20% B); T = 35.00 min (80% A, 20% B) gradient to T = 35.01 min (0% A, 100% B); T = 38.00 min (0% A, 100% B); gradient to T = 38.01 min (85% A, 15% B); T = 40.00 min (85% A, 15% B); Flow rate= 13ml / min; analysis time 40.00 min.Example 22: (E)- / V-(5-Chloro-2-(5-chloro-2-hvdroxy-4-methoxybenzoyl)isoindolin-4-yl)-N-methyl-3-(1- methylpyrrolidin-2-yl) acrylamideStep-1 : f-Butyl-(E)-5-chloro-4-(A / -methyl-3-(1-methylpyrrolidin-2-yl) acrylamido)isoindoline-2-carboxylate

[0433] Carried out 2 parallel batches at 0.4g scale: A stirred solution of f-butyl 5-chloro-4-(methylamino) isoindoline-2-carboxylate (0.4 g, 1.61 mmol, 1.0 eq, Example 15, Step 3) in DCM (5.0 mL) at room temperature was treated with (E)-3-(1-methylpyrrolidin-2-yl) acrylic acid (CAS 2400937-89-3, 0.49 g, 3.22 mmol, 2.0 eq) and pyridine (0.51 mL, 6.44 mmol, 4.0 eq). The mixture was cooled to 0°C and treated dropwise with POCh (0.22 mL, 2.41 mmol, 1.5 eq). The resulting reaction mixture was stirred at room temperature for 16h, then poured into water (100 mL), and extracted with DCM (3 x 100 mL).The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to give crude material (0.56 g), which was purified by flash column chromatography (product eluted in 4% MeOH in DCM) yielding f-butyl-(E)-5-chloro-4-(A / -methyl-3-(1-methylpyrrolidin-2-yl) acrylamido)isoindoline-2- carboxylate as a light brown solid (0.45g, Yield: 33%, 1.07 mmol).

[0434] 1H NMR (DMSO-d6, 400 MHz): 6 ppm 1 .45 (s, 9H), 1 .59 - 1 .60 (m, 1 H), 1 .79 (s, br, 2H), 1 .96 (s, br, 1 H), 2.29 (s, br, 2H), 2.85 - 2.94 (m, 1 H), 3.12 (s, 3H), 4.40 - 4.66 (m, 4H), 5.86 (s, br, 1 H), 6.57 - 6.66 (m, 1 H), 7.31 - 7.46 (m, 1 H), 7.57 - 7.60 (m, 1 H), 10.31 (bs, 1 H).

[0435] LCMS (Method N): 1.761 min, 1.777 min, MS: ES+ 420.2, 422.2 (M, M+2).Step-2: (E)- / V-(5-Chloroisoindolin-4-vl)- / V-methvl-3-(1-methvlpvrrolidin-2-vl)acrvlamide hydrochloride

[0436] A stirred solution of f-butyl-(E)-5-chloro-4-(A / -methyl-3-(1-methylpyrrolidin-2-yl) acrylamido)isoindoline-2-carboxylate (Step 1) (0.45 g, 1.07 mmol, 1.0 eq) in DCM (5.0 mL) was cooled to 0°C and treated dropwise with 4M HCI in dioxane (2.5 ml, 5v). After stirring for 1 hr, the reaction mixture was concentrated under high vacuum to give crude material (0.45 g), which was triturated using diethyl ether yielding (E)- / V-(5-chloroisoindolin-4-yl)- / V-methyl-3-(1 -methylpyrrolidin-2-yl)acrylamide hydrochloride as a navy blue solid (0.45 g, Yield: Quantitative, 1 .40 mmol).

[0437] 1H NMR (DMSO-d6, 400 MHz): 5 ppm 1.72 - 1.77 (m, 1 H), 1.92 - 1.94 (m, 2H), 2.13 - 2.18 (m, 1 H), 2.57 (d, J = 4.0 Hz, 2H), 2.94 - 2.95 (m, 1 H), 3.13 (s, 3H), 3.79 - 3.87 (m, 1 H), 4.42 - 4.57 (m, 4H), 6.02 - 6.09 (m, 1 H), 6.68 - 6.80 (m, 1 H), 7.49 - 7.52 (m, 1 H), 7.66 (d, J = 8.0Hz, 1 H), 10.10 (s, br, 1 H), 10.50 (s, br, 1 H), 10.99 (d, J = 27.2 Hz, 1 H).

[0438] LCMS (Method O: 6.842 min, MS: ES+ 320.3 (M+1).Step 3: (E)- / V-(5-Chloro-2-(5-chloro-2-hydroxy-4-methoxybenzoyl)isoindolin-4-yl)- / V-methyl-3-(1- methylpyrrolidin-2-yl)acrylamide

[0439] A stirred solution of 5-chloro-2-hydroxy-4-methoxybenzoic acid (CAS 1378866-39-7) (0.2 g, 0.98 mmol, 1.0 eq.) in DMF (2.0 mL) at room temperature was treated with EDC. HCI (0.28 g, 1.48 mmol, 1.5 eq) and HOAt (0.13 g, 0.98 mmol, 1.0 eq). (E)- / V-(5-chloroisoindolin-4-yl)-N-methyl-3-(1-methylpyrrolidin- 2-yl) acrylamide hydrochloride (Step 2) (0.47 g, 1 .48 mmol, 1 ,5eq) and NMM (0.54 mL, 4.93 mmol, 5.0 eq) were added to the reaction mixture which was stirred at room temperature for 16h , then poured into water (50 mL) and extracted with EtOAc (3 x 50 mL). The combined organic layer was dried over Na2SC>4 and concentrated under reduced pressure to give crude material (0.26 g), which was purified by reverse phase chromatography (product eluted at 30% acetonitrile in water) yielding (E)- / V-(5-chloro-2-(5-chloro-2-hydroxy-4-methoxybenzoyl)isoindolin-4-yl)-N-methyl-3-(1 -methylpyrrolidin-2-yl) acrylamide as a white solid (0.035 g, Yield: 16%, 0.069 mmol).

[0440] 1H NMR (DMSO-d6, 400 MHz, 349K): 6 ppm 1.27 - 1.32 (m, 1 H), 1.47 - 1.48 (m, 1 H), 1.71 (m, 2H), 1.87-1.93 (m, 1 H), 2.12 (s, br, 3H), 2.27 - 2.28 (m, 1 H), 2.80 (m, br, 1 H), 3.20 (s, 3H), 3.87 (s, 3H), 4.63 (d, J= 15.6 Hz, 1 H), 4.79 - 4.84 (m, 3H), 5.74 (d, J = 14.8 Hz, 1 H), 6.54 - 6.59 (m, 1 H), 6.70 (s, 1 H), 7.29 - 7.32 (m, 1 H), 7.43 (d, J = 8.0 Hz, 1 H), 7.57 (d, J = 8.0 Hz, 1 H), 10.41 (bs, 1 H).

[0441] LCMS (Method N): 1.735 min, 1.762 min, MS: ES+ 504.2, 506.2 (M, M+2).

[0442] HPLC (Method I): 4.487 min and 4.593 min.Preparation of IntermediatesIntermediate A: (E)-4-(Cvclopropyl(methyl)amino)- / V-(isoindolin-4-yl)but-2-enamide dihydrochlorideStep 1 : te / Y-Butyl (E)-4-(4-(cyclopropyl(methyl)amino)but-2-enamido)isoindoline-2-carboxylate

[0443] To a stirred solution of (E)-4-(cyclopropyl(methyl)amino)but-2-enoic acid (CAS-1418272-86-2) (410 mg, 2.6 mmol, 1.4 eq), HATU (876 mg, 2.3 mmol, 1.2 eq) and te / Y-butyl 4-aminoisoindoline-2- carboxylate (CAS 871013-98-8, 450 mg, 1.9 mmol, 1.0 eq) in DCM (10.00 mL) was added DIPEA (0.67 mL, 3.84 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 2 h then diluted with DCM (30 mL) and washed with water (30 mL). The aqueous phase was re-extracted with DCM (2 x 20 mL) and the combined organic phases were washed with brine (30 mL), filtered through a hydrophobic frit and concentrated under reduced pressure. The crude residue was purified by chromatography on silica eluting with 0-100% EtOAc in cyclohexane to afford the title compound (270 mg, Yield: 38%).

[0444] MS:ES+ 372.1 (M+1).

[0445] 1H NMR (400 MHz, CDCI3) 6, 0.52 - 0.42 (m, 4H), 1 .53 - 1 .52 (m, 9H), 1 .79 - 1 .73 (m, 1 H), 2.38 - 2.34 (m, 3H), 3.36 (d, J=4.5 Hz, 2H), 4.72 - 4.60 (m, 4H), 6.13 - 6.04 (m, 1 H), 6.84 (s, 1 H), 7.05 - 6.96 (m, 2H), 7.31 - 7.27 (m, 1 H), 7.89 - 7.74 (m, 1 H).Step 2: (E)-4-(Cyclopropyl(methyl)amino)- / V-(isoindolin-4-yl)but-2-enamide dihydrochloride

[0446] To a solution of te / Y-butyl (E)-4-(4-(cyclopropyl(methyl)amino)but-2-enamido)isoindoline-2- carboxylate (Step 1) (270 mg, 0.73 mmol, 1.0 eq) in DCM (5.0 mL) and MeOH (2.0 mL) was added 3M HCI in CPME (4.8 mL, 14.5 mmol, 20.0 eq). The reaction mixture was stirred at room temperature for 1 h under an inert atmosphere. The resulting mixture was concentrated under reduced pressure to afford the title compound (300 mg, Yield quantitative). The material was used in the next steps without further purification.

[0447] MS:ES+ 272.1 (M+1).Intermediate B: (E)-4-(Diethylamino)- / V-(isoindolin-4-yl)but-2-enamide dihydrochlorideStep 1 : Ethyl (E)-4-(diethylamino)but-2-enoate

[0448] A mixture of ethyl (E)-4-bromo-2-butenoate (CAS 37746-78-4; 0.78 mL, 5.7 mmol, 1.0 eq), diethylamine (1 .8 mL, 17 mmol, 3.0 eq) and potassium carbonate (1 .2 g, 8.6 mmol, 1 .5 eq) in THF (15 mL) was heated to 50°C in a pressure tube for 20 h. The resulting mixture was cooled to room temperature and filtered through Celite® . The filtrate was concentrated under reduced pressure to afford the title compound (1.1 g, quantitative) which was used in the next step without further purification.

[0449] MS:ES+ 186.1 (M+1).Step 2: (E)-4-(Diethylamino)but-2-enoic acidNaOH

[0450] A solution of ethyl (E)-4-(diethylamino)but-2-enoate (Step 1) (1.1 g, 5.7 mmol, 1.0 eq) and 2M NaOH (2.8 mL, 5.7 mmol, 1 .0 eq) in THF (10 mL) was heated to 50°C for 18 h. Additional 2M NaOH (0.28 mL, 0.57 mmol, 0.10 eq) was added and heating continued at 50°C for a further 7 h. The resulting mixture was allowed to cool to room temperature and stand overnight. The pH was adjusted to pH 6 with 2N HCI then the mixture was concentrated under reduced pressure. The residue was stirred with 10% MeOH in DCM (10 mL) for 2 h, filtered and concentrated under reduced pressure to give an orange oil which solidified on standing to afford the title compound (0.76 g, Yield: 85%).

[0451] MS:ES+ 158.0 (M+1).

[0452] 1H NMR (400 MHz, DMSO) 6 1 .00 - 0.96 (m, 6H), 2.57 - 2.47 (m, 4H), 3.29 - 3.23 (m, 2H), 5.99 - 5.93 (m, 1 H), 6.81 - 6.74 (m, 1 H).Step 3: (E)-4-(Diethylamino)but-2-enoyl chloride

[0453] Oxalyl chloride (0.072 mL, 0.83 mmol, 1.3 eq) was added to a suspension of (E)-4- (diethylamino)but-2-enoic acid (Step 2) (100 mg, 0.64 mmol, 1 .0 eq) in acetonitrile (6.5 mL) followed by 2 drops of DMF at 0°C. The resulting orange solution was stirred for 7 h then used in next step without further purification.Step 4: (E)-4-(Diethylamino)- / V-(isoindolin-4-yl)but-2-enamide dihydrochloridei) MeCN 2HCI ii) HCI, CPME

[0454] A solution of (E)-4-(diethylamino)but-2-enoyl chloride (Step 3) (112 mg, 0.636 mmol, 1.0 eq) in acetonitrile (6.5 mL) was added to a solution of tert-butyl 4-aminoisoindoline-2-carboxylate (CAS: 871013- 98-8) (150 mg, 0.64mmol, 1 .0 eq) in acetonitrile (6.5 mL) at 0°C. The reaction mixture was stirred for 15 h allowing the reaction to come to room temperature. The resulting mixture was concentrated under reduced pressure and the residue was dissolved in 3M HCI in CPME (5 mL) and MeOH (3 mL). The resulting solution was stirred for 4 h then concentrated under reduced pressure. The residue was dried under high vacuum overnight to afford the title compound as a cream solid which was used without further purification. (0.26 g, Yield: quantitative).

[0455] MS:ES+ 274.1 (M+1).Intermediate C: 3-Chloro-4,6-dihvdroxy-2-methylbenzoic acidStep 1 : 2,4-Dihydroxy-6-methylbenzaldehyde

[0456] A cooled (0 °C) solution of 5-methylbenzene-1 ,3-diol (3.0 g, 24.16 mmol, 1.0 eq.) (CAS: 504-15- 4) in DMF (30 mL) was treated dropwise with POCh (7.41 g, 48.33 mmol, 2.0 eq.). The reaction mixture was allowed to warm to room temperature and stirred for 16 h. The resulting mixture was poured into ice cold water (50 mL) and basified with 10% NaOH solution causing a suspension to form. The solid wascollected by filtration and dried under reduced pressure. The crude material was triturated with n-pentane (2 x 20 mL) followed by high vacuum drying to afford the title compound as an off-white solid (1.8 g, 44.0%).

[0457] 1H NMR (Acetone, 400 MHz): 6 2.55 (s, 3H), 6.18 (d, J= 4 Hz, 1 H), 6.31 (s, 1 H), 9.73 (s, 1 H, D2O exchangeable), 10.1 1 (d, J= 2 Hz, 1 H), 12.51 (s, 1 H, D2O exchangeable).

[0458] LCMS (Method A): 1.552 min, MS: ES+ 153.1 (M+1).Step 2: 3-Chloro-4,6-dihydroxy-2-methylbenzoic acid

[0459] A cooled (0 °C) solution of 2,4-dihydroxy-6-methylbenzaldehyde (0.5 g, 3.28 mmol, 1.0 eq.) in THF: H2O (2:1) was treated dropwise with sulfamic acid (0.63 g, 6.57 mmol, 2.0 eq.) and NaCIC>2 (0.59 g, 6.57 mmol, 2.0 eq.) and the reaction mixture was stirred at 0°C for 2 h. The resulting mixture was poured into water (10 mL) and extracted with ethyl acetate (3 x 15 mL). The combined organic extracts were dried over Na2SO4 and concentrated under reduced pressure. The crude material was triturated with n-pentane (3 x 10 mL) and dried under high vacuum to afford the title compound as a yellow solid (0.5 g, 75.32%).

[0460] 1H NMR (DMSO-d6, 400 MHz): 6 2.37 (s, 3H), 6.41 (s, 1 H), 10.56 (s, 1 H), 10.95 (s, 1 H, D2O exchangeable), 12.98 (s, 1 H, D2O exchangeable).Intermediate D: 3-Chloro-6-hvdroxy-4-methoxy-2-methylbenzoic acidStep 1 : (6-Chloro-7-hydroxy-2,2,5-trimethyl-4 / 7-benzo[d][1 ,3]dioxin-4-one

[0461] To a stirred solution 3-chloro-4,6-dihydroxy-2-methylbenzoic acid (Intermediate C) (2.0 g, 9.87 mmol, 1 .0 eq.) in DMF (20 mL) at room temperature were added acetone (7.3 mL, 29.6 mmol, 10 eq.) and DMAP (0.12 g, 0.98 mmol, 0.1 eq.). The mixture was cooled to 0°C and treated dropwise with thionyl chloride (2.15 mL, 29.61 mmol, 3 eq.). The reaction mixture was allowed to warm to room temperature and stirred for 2 h. The resulting mixture was neutralized with sat. NaHCCh (30 mL) and extracted with ethyl acetate (2 x 40 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude material was purified by chromatography on silica (product was eluted in 100% DCM) yielding (6-chloro-7-hydroxy-2,2,5-trimethyl-4 / 7-benzo[d][1 ,3]dioxin-4-one as a white solid (1.0 g, Yield: 41 %).

[0462] 1H NMR (DMSO-d6, 400 MHz): 6 1.64 (s, 6H), 2.65 (s, 3H), 6.49 (s, 1 H), 11.57 (s, 1 H).

[0463] LCMS (Method N): 2.14 min, MS: ES+ 242.78 (M+1).Step 2: 6-Chloro-7-methoxy-2,2,5-trimethyl-4H-benzo[d][1 ,3]dioxin-4-one

[0464] To a stirred solution of (6-chloro-7-hydroxy-2,2,5-trimethyl-4 / 7-benzo[d][1 ,3]dioxin-4-one (Step 1) (1 g, 4.12 mmol, 1.0 eq.) in acetonitrile (10 mL) was added K2CO3 (0.68g, 4.94 mmol, 1.2 eq.) and the mixture was stirred at room temperature for 10 mins. Dimethyl sulphate (0.54g, 4.32 mmol, 1 .05 eq.) was added and stirring continued at room temperature for 16 h. The resulting mixture was diluted with water (20 mL) and extracted with ethyl acetate (2 x 20 mL). The combined organic extracts were dried over Na2SC>4, filtered and concentrated under reduced pressure. The crude material was purified by chromatography on silica (product was eluted in 100% DCM) yielding 6-chloro-7-methoxy-2,2,5-trimethyl- 4 / 7-benzo[d][1 ,3]dioxin-4-one as a yellow gummy (0.8 g, Yield: 75%).

[0465] 1H NMR (DMSO-d6, 400 MHz): 6 1.67 (s, 6H), 2.68 (s, 3H), 3.92 (s, 3H), 6.78 (s, 1 H).Step 3: 3-Chloro-6-hydroxy-4-methoxy-2-methylbenzoic acid

[0466] To a stirred solution of 6-chloro-7-methoxy-2,2,5-trimethyl-4 / 7-benzo[d][1 ,3]dioxin-4-one (Step 2) (0.7 g, 2.73 mmol, 1.0 eq.) in THF:H2O (1 : 1) (14 mL) was added LiOH.H2O (1.14 g, 27.3 mmol, 10 eq.) and the reaction mixture was stirred at room temperature for 16 h. The resulting mixture was acidified with dilute HCI and extracted with ethyl acetate (2 x 30 mL). The combined organic extracts were dried over Na2SC>4, filtered, and concentrated under reduced pressure yielding 3-chloro-6-hydroxy-4-methoxy-2- methylbenzoic acid as a white solid (0.4 g, Yield: 76%).

[0467] 1H NMR (DMSO-d6, 400 MHz): 6 2.40 (s, 3H), 3.83 (s, 3H), 6.52 (s, 1 H), 12.35 (bs, 1 H).Intermediate E: / V-(6-(2-(Dimethylamino)ethoxy)isoindolin-4-yl)- / V-methylacrylamide. TFA saltStep 1 : 7-Bromoisoindolin-5-ol hydrochloride

[0468] BBrs (1 M in DCM) (40 mL) was added dropwise to 4-bromo-6-methoxyisoindoline hydrochloride (CAS 3050902-22-9) (2.0 g, 7.6 mmol, 1 eq.) at room temperature and heated to 60°C for 16 h. Theresulting mixture was concentrated under reduced pressure and the crude material was diluted with water (50 mL) causing a solid to precipitate. The solid was collected by filtration and dried under high vacuum to yield the title compound as an off-white solid (1 .35 g, Yield: 72. 0%).

[0469] 1H NMR (DMSO-d6, 400 MHz): 5 3.94 (s, 2H), 4.24 (s, 2H), 6.27 - 6.31 (m, 1 H), 6.42 - 6.46 (m, 1 H).

[0470] LCMS (Method A): 0.688 min, MS: ES+ 213.9 & 215.9 (M+1 & M+3).Step 2: te / Y-Butyl 4-bromo-6-hydroxyisoindoline-2-carboxylateBoc anhydride,

[0471] To a stirred solution of 7-bromoisoindolin-5-ol hydrochloride (Step 1) (1.3 g, 6.1 mmol, 1 eq.) in DCM (26 mL) was added TEA (1 .85 g, 18.3 mmol, 3 eq.) and the mixture was stirred at room temperature for 15 mins. Boc-anhydride (1 .33 g, 6.1 mmol, 1 eq.) was added and stirring continued at room temperature for 2 h. The resulting mixture was diluted with water (50 mL) and extracted with DCM (5 x 30 mL). The combined organic extracts were dried over Na2SC>4, filtered, and concentrated under reduced pressure. The crude material was purified by chromatography on silica (product eluted at 45% EtOAc in hexane) to yield the title compound as an off-white solid (1 .150 g, Yield: 55. 0%).

[0472] 1H NMR (DMSO-d6, 400 MHz): 6 1.45 (s, 9H), 4.39 (d, J= 10.4 Hz, 2H), 4.59 (d, J= 10.4 Hz, 2H), 6.73 (s, 1 H), 6.87 (d, J= 1 .6 Hz, 1 H), 9.91 (s, 1 H).

[0473] LCMS (Method A): 2.024 min, MS: ES+ 257.8 (M-56).Step 3: 4-Bromo-6-(2-(dimethylamino)ethoxy)isoindoline-2-carboxylate

[0474] To a stirred solution of te / Y-butyl 4-bromo-6-hydroxyisoindoline-2-carboxylate (Step 2) (0.2 g, 0.63 mmol, 1 eq.) in THF (10 mL) was added KOtBu (0.353 g, 3.15 mmol, 5 eq.) and the mixture was stirred at room temperature for 1 h. 2-Bromo- / V, / V-dimethylethan-1 -amine hydrobromide (CAS 2862-39-7) (0.447 g, 1 .91 mmol, 3 eq.) was added and the reaction mixture was heated to 70°C for 6 h. The resulting mixture was diluted with water (100 mL) and extracted with EtOAc (4 x 30 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude material was triturated with diethyl ether (2 x 15 mL) and dried under high vacuum to yield the title compound as a yellow solid (1 .05 g, Yield: 63.0%).

[0475] 1H NMR (DMSO-d6, 400 MHz): 6 1.45 (s, 9H), 2.19 (s, 6H), 2.59 (d, J= 5.6 Hz, 2H), 4.03 - 4.06 (m, 2H), 4.44 (d, J= 10.4 Hz, 2H), 4.64 (d, J= 6.0 Hz, 2H), 6.96 (d, J= 6.8 Hz, 1 H), 7.09 (bs, 1 H).

[0476] LCMS (Method A): 1.461 min, MS: ES+ 384.9 & 386.9 (M+1 & M+3).Step 4: fe / Y-Butyl-6-(2-(dimethylamino)ethoxy)-4-(methylamino)isoindoline-2-carboxylateCH2NH2(2M) NaOtBu, tBu-Xphos

[0477] Performed in 5 parallel batches, each of 0.4 g scale: To a stirred solution of tert-butyl 4-bromo- 6-(2-(dimethylamino)ethoxy)isoindoline-2-carboxylate (Step 3) (0.4 g, 1.03 mmol, 1.0 eq.) in THF (4.0 mL) was added NaOtBu (0.19 g, 2.07 mmol, 2.0 eq) at room temperature and the mixture was purged with argon for 20 mins. Methylamine (2M in THF) (1 mL, 2.07 mmol, 2.0 eq) and f-Bu-XphosPdG3 (0.049 g, 0.062 mmol, 0.06 eq) were added and the reaction mixture was heated to 70°C temperature for 30 mins. The resulting mixture was cooled to room temperature, diluted with water (50 mL) and extracted with EtOAc (3 x 50 mL). The combined organic extracts were dried over Na2SC>4, filtered, and concentrated under reduced pressure. The crude material was purified by chromatography on silica (product eluted at 6% MeOH in DCM) yielding tert-butyl-6-(2-(dimethylamino)ethoxy)-4-(methylamino)isoindoline-2-carboxylate as a yellow solid (1 .2 g, Yield: 69%).

[0478] 1H NMR (DMSO-d6, 400 MHz): 6 1.45 (d, J= 3.6 Hz, 9H), 2.21 (d, J= 8.8 Hz, 6H), 2.58 (t, J= 6 Hz, 2H), 2.66 (t, J= 4.4 Hz, 3H), 3.96 (t, J= 5.2 Hz, 2H), 4.29 (d, J= 10.4 Hz, 2H), 4.44 (d, J= 9.6 Hz, 2H), 5.45 (t, J= 5.2 Hz, 1 H, D2O exchangeable), 5.90 (s, 1 H), 6.12 (d, J= 7.2 Hz, 1 H).

[0479] LCMS (Method N): 1.805 min, MS: ES+ 335.9 (M+1).Step 5: te / Y-Butyl 6-(2-(dimethylamino)ethoxy)-4-( / V-methylacrylamido)isoindoline-2-carboxylate

[0480] To a cooled (0°C) solution of tert-butyl 6-(2-(dimethylamino)ethoxy)-4-(methylamino) isoindoline- 2-carboxylate (Step 4) (0.2 g, 0.59 mmol, 1 .0 eq.) in DCM (2 mL) was added TEA (0.42 g, 4.17 mmol, 7.0 eq.) and the mixture was allowed to stir for 10 mins. Acryloyl chloride (0.27 g, 2.98 mmol, 5.0 eq.) was added and stirring continued at 0°C for 15 mins. The resulting mixture was diluted with cold water (30 mL) and extracted with DCM (3 x 30 mL). The combined organic extracts were dried over Na2SO4, filtered and concentrated under reduced pressure. The crude material was triturated with n-pentane (2 x 10 mL) yielding tert-butyl 6-(2-(dimethylamino)ethoxy)-4-( / V-methylacrylamido)isoindoline-2-carboxylate as yellow liquid (0.35 g, Yield: Quantitative). The crude material was used without further purification.

[0481] LCMS (Method N): 1.644 min, MS ES+: 390.4 (M+1).Step 6: / V-(6-(2-(Dimethylamino)ethoxy)isoindolin-4-yl)- / V-methylacrylamide. TFA salt

[0482] To a cooled (0°C) solution of fe / Y-butyl 6-(2-(dimethylamino)ethoxy)-4-( / V-methylacrylamido) isoindoline-2-carboxylate (Step 5) (0.3 g, 0.77 mmol, 1 .0 eq) in DCM (3 mL) was added TFA (1 .5 mL) and the reaction mixture was stirred at room temperature for 30 mins. The resulting mixture was concentrated under reduced pressure and the crude material was triturated with n-pentane (2 x 10 mL) yielding fe / Y-butyl 6-((dimethylamino)ethoxy)-4-( / V-methylacrylamido)isoindoline-2-carboxylate 2,2,2-trifluoroacetate as a brown liquid (0.45 g, Yield: quantitative) which was used directly.

[0483] LCMS (Method O): 1.315 min, MS ES+: 290.1 (M+1).Intermediate F: 6-Hvdroxy-4-methoxy-2,3-dimethylbenzoic acidStep 1 : 4,5-Dimethyl-1 ,3-phenylene diacetate

[0484] Performed in 2 parallel batches, each of 30 g scale: A stirred solution of 5,5- dimethylcyclohexane-1 ,3-dione (CAS: 126-81-8) (30 g, 214.5 mmol, 1.0 eq.) in acetic anhydride (340 mL) at room temperature, was treated dropwise with concentrated H2SO4 (12.4 mL, 235 mmol, 1.1 eq.). The reaction mixture was heated to 150°C and stirred for 1 h. The resulting mixture was slowly poured into ice cold water (500 mL) and extracted with ethyl acetate (3 x 500 mL). The combined organic extracts were washed with NaHCCh solution (3 x 500 mL), dried over Na2SC>4 and concentrated under reduced pressure. The crude material was purified by chromatography on silica (product was eluted in 15% EtOAc in hexane) yielding the title compound as a white solid (80 g, Yield: 84%).

[0485] 1H NMR (DMSO-d6, 400 MHz): 6 2.24 (s, 3H), 2.26 (s, 3H), 2.30 (s, 3H), 2.32 (s, 3H), 6.76 (d, J = 2.4 Hz, 1 H), 6.88 (d, J = 2.4 Hz, 1 H).Step 2: 4, 5-Dimethylbenzene-1 ,3-diol

[0486] To a solution of 4, 5-dimethyl-1 ,3-phenylene diacetate (Step 1) (80 g, 360.3 mmol, 1.0 eq) in EtOH; H2O (1 :1) (800 mL) was added NaOH (57.6 g, 1441 .4 mmol., 4 eq.) and the mixture heated to 70°Cfor 2 h. The resulting mixture was acidified to pH 5 with dilute HCI and concentrated under reduced pressure. The crude material was diluted with ethyl acetate (300 mL), washed with brine solution (3 x 300 mL), dried over Na2SC>4 and concentrated under reduced pressure. Purification by chromatography on silica (product was eluted in 15% EtOAc in hexane) afforded the title compound (25 g, Yield: 55%).

[0487] 1H NMR (DMSO-d6, 400 MHz): 6 1.83 (s, 3H), 2.06 (s, 3H), 6.03 (s, 1 H), 6.12 (s, 1 H), 8.78 (s, 1 H) 8.94 (s, 1 H).Step 3: 4,6-Dihydroxy-2,3-dimethylbenzaldehyde

[0488] Phosphorus oxychloride (41 .5 g, 271 .7 mmol, 1 .5 eq.) was added dropwise to DMF (250 mL) at 0°C. To this solution was added dropwise 4,5-dimethylbenzene-1 ,3-diol (Step 2) (25 g, 181 .1 mmol, 1 eq.) in DMF at 0 °C and the reaction mixture was stirred at room temperature for 1 h. The resulting mixture was slowly poured into ice cold water, basified using NaOH solution (pH ~10) and acidified with dilute HCI (pH 4 - 5) causing precipitation of a solid. The solid was collected by filtration and dried under high vacuum to afford the title compound as a yellow solid (15 g, Yield: 65%).

[0489] 1H NMR (DMSO-d6, 400 MHz): 6 1.99 (s, 3H), 2.41 (s, 3H), 6.22 (s, 1 H), 10.13 (s, 1 H), 10.75 (s, 1 H) 12.24 (s, 1 H).

[0490] LCMS (Method A): 1.467 min, MS: ES+ 166.8 (M+1).Step 4: 4,6-Dihydroxy-2,3-dimethylbenzoic acid

[0491] A solution of 4,6-dihydroxy-2,3-dimethylbenzaldehyde (Step 3) (5 g, 30.12 mmol, 1.0 eq.) in DMSO: water (50 mL) was treated portionwise with NaCIC>2 (13.5 g, 150.6 mmol, 5 eq.) followed by NaH2PC>4 (18.0 g, 150.6 mmol, 5 eq.) and the reaction mixture stirred at room temperature for 1 h. The resulting mixture was diluted with water (100 mL) and acidified to pH 4-5 with dilute HCI. The mixture was extracted with ethyl acetate (3 x 80 mL) and the combined organic extracts were washed with brine solution (3 x 100 mL), dried over Na2SC>4 and concentrated under reduced pressure to afford the title compound (25 g, Yield: 55%).

[0492] 1H NMR (DMSO-d6, 400 MHz): 6 1.96 (s, 3H), 2.27 (s, 3H), 6.24 (s, 1 H), 9.86 (s, 1 H), 10.89 (s, 1 H) 13.07 (bs, 1 H).

[0493] LCMS (Method A): 1.220 min, MS: ES+ 182.9 (M+1).Step 5: 7-Hydroxy-2,2,5,6-tetramethyl-4H-benzo[d][1 ,3]dioxin-4-one

[0494] Performed in 5 parallel batches, each of 0.55 g scale: To a stirred solution of 4,6-dihydroxy-2,3- dimethylbenzoic acid (Step 4) (0.55 g, 3.02 mmol, 1.0 eq.) in DME (5.5 mL) were added SOCh (0.97 mL, 13.59 mmol, 4.5 eq.) and DMAP (0.055 g, 0.45 mmol, 0.15 eq.) at 0°C. The mixture was stirred for 5 mins at 0°C then treated with acetone (2.24 mL, 30.21 mmol, 10.0 eq.). The reaction mixture was allowed to warm to room temperature and stirred for 2 h. The resulting mixture was poured into cold saturated NaHCCh solution (500 mL) and extracted with ethyl acetate (2 x 200 mL). The combined organic extracts were dried over Na2SC>4, filtered and concentrated under reduced pressure. The crude material was purified by chromatography on silica (product eluted in 15% EtOAc in hexane) to yield 7-hydroxy-2, 2,5,6- tetramethyl-4H-benzo[d][1 ,3]dioxin-4-one as a yellow solid (0.28 g, Yield: 8%).

[0495] 1H NMR (DMSO-d6, 400 MHz): 6 1.60 (s, 6H), 2.09 (s, 3H), 2.50 (s, 3H), 6.32 (s, 1 H), 10.66 (s, 1 H).

[0496] LCMS (Method N): 1.884 min, MS ES+: 223.1 (M+1).Step 6: 7-Methoxy-2,2,5,6-tetramethyl-4 / 7-benzo[d][1 ,3]dioxin-4-one

[0497] To a stirred solution of 7-hydroxy-2,2,5,6-tetramethyl-4H-benzo[d][1 ,3]dioxin-4-one (Step 5) (0.28 g, 1.26 mmol, 1.0 eq.) in DMF (2.8 mL) was added K2CO3 (0.522 g, 3.78 mmol, 3.0 eq.) and the mixture was stirred for 10 mins at room temperature. The mixture was cooled to 0°C, treated dropwise with Mel (3.58 mL, 2.52 mmol, 2.0 eq) and gradually allowed to warm to room temperature stirring for 3 h. The resulting mixture poured into ice cold water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic phase dried over Na2SO4, filtered and concentrated under reduced pressure. The crude material was purified by chromatography on silica (product eluted in 7% EtOAc in hexane). The product fractions were combined and concentrated under reduced pressure to yield 7-methoxy-2,2,5,6-tetramethyl-4H- benzo[d][1 ,3]dioxin-4-one as a yellow crystalline solid (0.17 g, 41 % yield).

[0498] 1H NMR (DMSO-d6, 400 MHz): 6 1.64 (s, 6H), 2.06 (s, 3H), 2.53 (s, 3H), 3.84 (s, 3H), 6.56 (s, 1 H).

[0499] LCMS (Method N): 2.219 min, MS ES+: 237.1 (M+1).Step 7: 6-Hydroxy-4-methoxy-2,3-dimethylbenzoic acid

[0500] To a stirred solution of 7-methoxy-2,2,5,6-tetramethyl-4H-benzo[d][1 ,3]dioxin-4-one (Step 6) (0.17 g, 0.72 mmol, 1 .0 eq.) in THF: water (1 :1) (3.4 mL) was added KOH (0.40 g, 7.20 mmol, 5.0 eq.) and the reaction mixture was heated to 70°C for 12 h. The resulting mixture was poured into water (10 mL) and washed with EtOAc (20 mL). Then aqueous portion was acidified with 1 N HCI (2 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over Na2SO4, filtered and concentrated under reduced pressure to yield 6-hydroxy-4-methoxy-2,3-dimethylbenzoic acid as a yellow solid (0.04 g, 28% yield).

[0501] 1H NMR (DMSO-d6, 400 MHz): 6 2.00 (s, 3H), 2.29 (s, 3H), 3.76 (s, 3H), 6.36 (s, 1 H), 11 .04 (bs, 1 H), 13.14 (bs, 1 H).

[0502] LCMS (Method N): 1.924 min, MS ES+: 195.2 (M-1).Intermediate G: (E)-4-(((Benzyloxy)carbonyl)(methyl)amino)but-2-enoic acidStep 1 : Methyl-(E)-4-(methylamino) but-2-enoate

[0503] Performed in 5 parallel batches, each of 5 g scale: To a cooled (-60°C) solution of methyl (E)-4- bromobut-2-enoate (CAS: 6000-00-6) (5 g, 27.93 mmol, 1.0 eq.) in THF (50 mL) was added dropwise methylamine (2M in THF) (35 mL) and the reaction mixture was stirred at room temperature for 4 h. The resulting mixture was diluted with THF (100 mL) and concentrated under reduced pressure. The crude material was purified by chromatography on silica (product eluted at 8% MeOH in DCM) yielding methyl (E)-4-(methylamino) but-2-enoate as a yellow liquid (8.0 g, Yield: 45%).

[0504] 1H NMR (DMSO-d6, 400 MHz): 6 2.5s (s, 3H), 3.70 (s, 3H), 3.79 (d, J= 6.4 Hz, 2H), 6.23 (d, J= 16 Hz, 1 H), 6.78 - 6.86 (m, 1 H), 8.74 (bs, 1 H).Step 2: Methyl (E)-4-(((benzyloxy) carbonyl) (methyl)amino)but-2-enoateCbz-CI K CO

[0505] Performed in 4 parallel batches, each of 2 g scale: To a stirred solution of methyl (E)-4- (methylamino)but-2-enoate (Step 1) (2.0 g, 15.48 mmol, 1.0 eq.) in DCM (20 mL) was added potassium carbonate (3.21 g, 23.20 mmol, 1.5 eq.) and the mixture was stirred at room temperature for 15 mins. Benzyl chloroformate (3.02 mL, 17.00 mmol, 1.1 eq.) was added dropwise and stirring continued at room temperature for 2 h. The resulting mixture was diluted with cold water (1000 mL) and extracted with DCM(2 x 1000 mL). The combined organic extracts were dried over Na2SC>4, filtered and concentrated under reduced pressure. The crude material was purified by chromatography on silica (product eluted at 20% EtOAc: hexane) yielding methyl-(E)-4-(((benzyloxy) carbonyl)(methyl)amino)but-2-enoate as yellow liquid (6.0 g, Yield: 36%). The material was used in the next step without further purification.Step 3: (E)-4-(((Benzyloxy)carbonyl)(methyl)amino)but-2-enoic acid

[0506] Performed in 2 parallel batches, each of 3.0 g scale: To a stirred solution of methyl (E)-4- (((benzyloxy) carbonyl) (methyl)amino)but-2-enoate (Step 2) (3.0 g, 11 .40 mmol, 1 .0 eq) in MeOH (30 mL) and water (30 mL) was added LiOH.H2O (2.39 g, 57.03 mmol, 5 eq.) and the reaction mixture was stirred at room temperature for 2 h. The resulting mixture was diluted with KHSO4 solution (100 mL) and extracted with EtOAc (3 x 200 mL). The combined organic extracts were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure yielding (E)-4-(((benzyloxy)carbonyl)(methyl)amino)but-2-enoic acid as yellow liquid (4.2 g, Yield: 74%). The material was used in the next step without further purification.Intermediate H: te / Y-Butyl-4-(methylamino)isoindoline-2-carboxylate2M Methyl amine in THF NaOtBu, Pd2(dba)3, Brettphos, 1 ,4-dioxane, 120°C, 1 h, MicrowaveBr HN^

[0507] Performed in 10 parallel batches, each of 1 g scale: To a stirred solution of fe / Y-butyl-4- bromoisoindoline-2-carboxylate (CAS:1035235-27-8) (1.0 g, 3.35 mmol, 1 eq.) in 1 ,4-dioxane (10 mL) was added 2M methyl amine in THF (2.5 mL, 5.03 mmol, 1 .5 eq) and NaO'Bu (0.806 g, 8.38 mmol, 2.5 eq) at room temperature. The mixture was degassed under nitrogen for 15 mins. Brettphos (0.108 g, 0.20 mmol, 0.06 eq) and Pd2(dba)3 (0.122 g, 0.13 mmol, 0.04 eq) were added and the reaction mixture was heated to 120°C using microwave irradiation for 1 h. The resulting mixture was filtered through Celite® and the filter cake washed with 10% MeOH in DCM (100 mL). The organic filtrate was concentrated under reduced pressure and purification of the crude material by chromatography on silica eluting with 15% EtOAc in hexane afforded the title compound as a light yellow solid (7.0 g, Yield: 84%).

[0508] 1H NMR (DMSO-d6, 400 MHz): 6 1.46 (s, 9H), 2.68 - 2.0 (m, 3H), 4.37 (d, J= 10 Hz, 2H), 4.50 (d, J= 10.4 Hz, 2H), 5.42 (dd, J= 4.8, 8.8 Hz, 1 H), 6.37 (d, J= 8.0 Hz, 1 H), 6.51 (t, J= 7.2 Hz, 1 H), 7.09 (t, J= 8.0 Hz, 1 H).

[0509] LCMS (Method A): 2.012 min, MS: ES+ 192.9 (M+1).Intermediate I: 2-Hvdroxy-5-isopropyl-4-methoxybenzoic acidStep 1 : Methyl 2,4-dihydroxybenzoate

[0510] To a stirred solution of 2,4-dihydroxybenzoic acid (CAS 89-86-1 , 100 g, 649.35 mmol) in MeOH (400 mL) was added concentrated H2SO4 (50 mL) at room temperature. The reaction mixture was heated to 100°C and stirred for 12 h. The resulting solution was concentrated under reduced pressure and poured into water (250 mL) causing a precipitate to form. The solids were collected by filtration and dried under high vacuum to afford the title compound as an off-white solid (85 g, 78%). This material was used in the next step without any further purification.

[0511] 1H NMR (CDCh, 400 MHz) 6 3.39 (s, 3H), 5.72 (s, br, 1 H), 6.38 - 6.42 (m, 2H), 7.74 (d, J= 8.8 Hz, 1 H), 11.02 (s, 1 H).

[0512] LCMS: 4.24 min, MS: ES+ 169. 19(M+1).Step 2: Methyl 2,4-dihydroxy-5-isopropylbenzoate

[0513] To a solution of methyl 2,4-dihydroxybenzoate (Step 1) (80 g, 475.7 mmol) in DCM (1200 mL) was added anhydrous AICI3 (126.87 g, 951 .48 mmol) followed by isopropyl bromide (351 .1 g, 2854.6 mmol) in 3 equal portions (117.0 g added at 6 h intervals) at 50 "C. The reaction mixture was stirred for a further 6 h at 50°C (total time 24 h) and then concentrated under reduced pressure. The crude material was diluted with water (1000 mL) and extracted with EtOAc (4 x 500 mL). The combined organic extracts were dried over Na2SC>4 and concentrated under reduced pressure. The resulting crude material was purified three times by chromatography on silica eluting with 3% EtOAc in hexane to afford the title compound as a yellow solid (25.5g, 26%).

[0514] 1H NMR (DMSO, 400 MHz) 6 1.13 (d, J= 7.6Hz, 6H), 3.03 - 3.16 (m, 1 H), 3.83 (s, 3H), 6.35 (s, 1 H), 7.49 (s, 1 H), 10.48 (s, 1 H), 10.57 (s, 1 H).

[0515] LCMS: 2.069 min, MS: ES+ 211.11 (M+1).Step 3: Methyl 2,4-dihydroxy-5-isopropylbenzoate

[0516] To a stirred solution of methyl 2,4-dihydroxy-5-isopropylbenzoate (Step 2) (0.7 g, 3.34 mmol, 1 .0 eq) in acetonitrile (7 mL) was added anhydrous K2CO3 (0.55 g, 3.9 mmol., 1.2 eq.) and the mixture was stirred at room temperature for 10 mins. Dimethyl sulphate (0.44 g, 3.5 mmol., 1.05 eq.) was added andstirring continued at room temperature for 16 h. The resulting mixture concentrated under reduced pressure and the crude material was diluted with water (20 mL) and extracted with EtOAc (2 x 100 mL). The combined organic extracts were dried over Na2SC>4, filtered, and concentrated under reduced pressure. The crude material was purified by chromatography on silica (product eluted in 5% EtOAc in hexane) yielding methyl 2-hydroxy-5-isopropyl-4-methoxybenzoate as an off-white solid (0.56 g, Yield: 75%).

[0517] 1H NMR (DMSO-d6, 400 MHz): 6 1.13 (d, J= 6.8 Hz, 6H), 3.07 - 3.14 (m, 1 H), 3.85 (s, 3H), 3.88 (s, 3H), 6.54 (s, 1 H), 7.54 (s, 1 H), 10.71 (s, 1 H).

[0518] LCMS (Method N): 2.731 min, MS: ES+ 225.05 (M+1).Step 4: 2-Hydroxy-5-isopropyl-4-methoxybenzoic acidOH O OH O1 11 KOH, MeOH JL K[| ^] ° Water, 70°C, 2h |l T °HMeO^y MeO^^y

[0519] To a stirred solution of methyl 2-hydroxy-5-isopropyl-4-methoxybenzoate (Step 3) (0.56 g, 2.5 mmol, 1.0 eq) in MeOH: H2O (5.6 mL) was added KOH (1.4 g, 2.5 mmol, 10 eq) and the reaction mixture was heated to 70°C for 2 h. The resulting mixture was poured into ice cold sat. solution of KHSO4 solution (10 mL) (pH adjusted to pH 5-6) and extracted with EtOAc (3 x 100 mL). The combined organic extracts were dried over Na2SO4, filtered and concentrated under reduced pressure yielding 2-hydroxy-5-isopropyl- 4-methoxybenzoic acid as a light pink solid (0.54 g, Yield: 99.51 %). The crude material was used without further purification.

[0520] 1H NMR (DMSO-d6, 400 MHz): 6 1.10 (d, J= 6.8 Hz, 6H), 3.05 - 3.09 (m, 1 H), 3.74 (s, 3H), 6.24 (s, 1 H), 7.46 (s, 1 H), 12.08 (bs, 1 H), 14.71 (bs, 1 H).

[0521] LCMS (Method N): 2.586 min, MS: ES+ 209.07 (M-1).Intermediate J: (E)-4-(Dimethylamino)- / V-methyl- / V-(3-methyl-1 ,2,3,4-tetrahydroisoquinolin-7-yl) but-2- enamide hydrochlorideStep 1 : 1 -(4-Bromophenyl)propan-2-amineTitanium tetraisopropoxide, 7N NH i M OH t 16h

[0522] To a stirred solution of 1-(4-bromophenyl)propan-2-one (CAS: 6186-22-7) (5.0 g, 23.4 mmol, 1 .0 eq.) in titanium tetra-isopropoxide (13.33 g, 2 eq) was added dropwise 7M ammonia in MeOH (60 mL) and the mixture was stirred at room temperature for 16 h. The mixture was cooled to 0°C, treated portion wise with NaBH4 (1.33 g, 35.12 mmol, 1.5 eq) and allowed to warm to room temperature stirring for 2 h. The resulting mixture was diluted with aqueous ammonia solution causing precipitation of a white solid. Thesolid was filtered and washed with EtOAc (200 mL). The filtrate was concentrated under reduced pressure causing further solid to precipitate out. EtOAc (200 mL) was added and the mixture was filtered. The filtrate was acidified with 2M HCI (200 mL) and the layers were separated. The pH of the aqueous layer was adjusted pH ~10 with aqueous ammonia (50 mL) and the mixture was extracted with DCM (500 mL). The organic extract was dried over Na2SO4, filtered and concentrated under reduced pressure yielding 1-(4- bromophenyl)propan-2-amine as a yellow solid (2.4 g, Yield: 48%). The material was used in the next step without further purification.

[0523] 1H NMR (DMSO-d6, 400 MHz): 5 0.93 (d, J= 6.4 Hz, 3H), 1.38 (s, br, 2H, D2O exchangeable), 2.50 (d, J= 3.6 Hz, 2H), 2.94 - 2.99 (m, 1 H), 7.14 (d, J= 7.6 Hz, 2H), 7.45 (d, J= 1 .2 Hz, 2H).

[0524] LCMS (Method N): 1.65 min, MS ES+: 216.2 (M).Step 2: Methyl(1-(4-bromophenyl) propan-2-yl)carbamatePyridine, Methyl

[0525] To a stirred solution of 1-(4-bromophenyl) propan-2-amine (Step 1) (2.4 g, 11.2 mmol, 1.0 eq.) in DCM (24 mL) was added dropwise pyridine (2.08 g, 26.35 mmol, 2.35 eq) and methylchloroformate (1.85 g, 19.02 mmol, 1 .75 eq) and the reaction mixture stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure. The crude material was dissolved in EtOAc (200 mL) and washed with sat. NaHCOs solution (200 mL), 1 N HCI (50 mL), brine solution (50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to yield methyl (1-(4-bromophenyl)propan-2- yl)carbamate as yellow solid (2.0 g, Yield: 48%). The material was used in the next step without further purification.

[0526] 1H NMR (DMSO-d6, 400 MHz): 6 1 .03 (d, J= 6.8 Hz, 3H), 2.56 - 2.69 (m, 2H), 3.46 (s, 3H), 3.65 - 3.68 (m, 1 H), 7.10 - 7.15 (m, 3H),7.45 (d, J= 8.4 Hz, 2H).Step 3: 7-Bromo-3-methyl-3,4-dihydroisoquinoline-2(1 H)-carboxylateParaformaldehyde, conc.H O rt, 16h

[0527] A mixture of concentrated H2SC>4 (11.13 mL) and glacial acetic acid (16.80 mL) were stirred for 15 mins then treated with methyl(1-(4-bromophenyl)propan-2-yl) carbamate (Step 2) (2.10 g, 7.71 mmol, 1.0 eq.) and paraformaldehyde (0.37 g, 12.34 mmol, 1 .60 eq.). The reaction mixture was stirred at room temperature for 16 h. The resulting mixture was diluted with ice cold water (500 mL) and extracted with DCM (2 x 50 mL). The combined organic extracts were dried over Na2SC>4, filtered and concentrated under reduced pressure. The crude material was triturated with diethyl ether (2 x 50 mL) and dried under high vacuum drying to yield methyl 7-bromo-3-methyl-3,4-dihydroisoquinoline-2(1 H)-carboxylate as a white solid (1.8 g, Yield: 87%).

[0528] 1H NMR (DMSO-d6, 400 MHz): 5 0.99 (d, J= 6.8 Hz, 3H), 2.58 - 2.62 (m, 1 H), 2.92 - 2.97 (m, 1 H), 3.64 (s, 3H), 4.27 (d, J= 17.2 Hz, 1 H), 4.46 (bs, 1 H), 4.69 (d, J= 17.2 Hz, 1 H), 7.15 (d, J= 8.4 Hz, 1 H), 7.36 - 7.39 (m, 1 H), 7.46 (s, 1 H).Step 4: 7-Bromo-3-methyl-1 ,2,3,4-tetrahydroisoquinoline lodotrimethylsilane,9 DCM M OH

[0529] To a stirred solution of methyl-7-bromo-3-methyl-3,4-dihydroisoquinoline-2(1 H)-carboxylate (Step 3) (2.0 g, 7.04 mmol, 1 .0 eq.) in DCM (40 mL) was added dropwise iodotrimethylsilane (5.63 g, 28.16 mmol, 4 eq) at room temperature. The reaction mixture was heated to 50°C and stirred for 2 h. The resulting mixture was diluted with MeOH (20 mL) and stirred for 10 mins. The mixture was concentrated under reduced pressure and the crude material was triturated with n-pentane (2 x 20 mL) and diethyl ether (5 x 20 mL) to yield 7-bromo-3-methyl-1 ,2,3,4-tetrahydroisoquinoline as a yellow solid (1.8 g, Yield: quantitative).

[0530] 1H NMR (DMSO-d6, 400 MHz): 6 1.34 (d, J= 6.4 Hz, 3H), 2.69 - 2.76 (m, 1 H), 3.01 - 3.07 (m, 1 H), 3.54 - 3.60 (m, 1 H), 4.35 (s, 2H), 7.19 (d, J= 18.0 Hz,1 H), 7.40 - 7.51 (m, 2H), 8.97 (bs, 1 H).Step 5: 7-Bromo-3-methyl-3,4-dihydroisoquinoline-2(1 H)-carboxylate Boc anhydride, TEA, PCM, rt, 1 hBr\xx^1^^N'Boc

[0531] To a cooled (0°C) solution of 7-bromo-3-methyl-1 ,2,3,4-tetrahydroisoquinoline (Step 4) (1.8 g, 7.96 mmol, 1 .0 eq.) in DCM (18 mL) were added dropwise TEA (3.32 mL, 23.89 mmol, 3 eq) followed by Boc anhydride (2.6 g, 11.9 mmol, 1 .5 eq). The reaction mixture was allowed to warm to room temperature and stirred for 1 h. The resulting mixture was diluted with ice cold water (50 mL) and extracted with DCM (3 x 50 mL). The combined organic extracts were dried over Na2SC>4, filtered and concentrated under reduced pressure to yield fe / Y-butyl-7-bromo-3-methyl-3,4-dihydroisoquinoline-2(1 H)-carboxylate as a yellow liquid (1 .7 g, Yield: 65%). The crude material was used in the next step without further purification.

[0532] 1H NMR (DMSO-d6, 400 MHz): 6 0.98 (d, J = 6.8Hz, 3H), 1 .43 (s, 9H), 2.50 - 2.60 (m, 1 H), 2.89 - 2.95 (m, 1 H), 4.22 (d, J= 18.4 Hz, 1 H), 4.41 (bs, 1 H), 4.63 (d, J= 17.2 Hz, 1 H), 7.13 (d, J= 8.0 Hz,1 H), 7.36 (d, J= 8.0 Hz, 1 H), 7.43 (s, 1 H).

[0533] LCMS (Method N): 2.260 min, MS ES+: 227.6 (M-100).Step 6: fe / Y-Butyl-3-methyl-7-(methylamino)-3,4-dihydroisoquinoline-2(1 H)-carboxylateMethylamine (40% in water), NaOtBu, f-But lX hosPdG3

[0534] The title compound was prepared from fe / Y-butyl-7-bromo-3-methyl-3,4-dihydroisoquinoline- 2(1 / - / )-carboxylate (Step 5) and methyl amine (40% in water) analogous to Intermediate E, Step 4.

[0535] 1H NMR (DMSO-d6, 400 MHz): 6 0.98 (d, J = 6.4 Hz, 3H), 1 .42 (s, 9H), 2.38 - 2.43 (m, 1 H), 2.64 (d, J= 5.2 Hz, 3H), 2.81 - 2.86 (m, 1 H), 4.10 (d, J= 5.6 Hz, 1 H), 4.35 (s, br, 1 H), 4.50 (d, J= 16.4 Hz, 1 H), 5.45 (d, J= 5.2 Hz,1 H, D2O exchangeable), 6.31 (s, 1 H), 6.38 - 6.41 (m, 1 H), 6.86 (d, J= 8.0 Hz, 1 H).

[0536] LCMS (Method N): 1.89 min, MS ES+: 221.15 (M-55).Step 7: fe / Y-Butyl-(E)-7-(4-(dimethylamino)- / V-methylbut-2-enamido)-3-methyl-3,4-dihydroisoquinoline-2(1 / - / )-carboxylatepyr ne,3DCM, O°C-rt, 2h

[0537] The title compound was prepared from (E)-4-(dimethylamino)but-2-enoic acid (CAS: 848133-35- 7) and fe / Y-butyl-3-methyl-7-(methylamino)-3,4-dihydroisoquinoline-2(1 H)-carboxylate (Step 6) analogously to Example 6, Step 1 .

[0538] 1H NMR (DMSO-d6, 400 MHz, D2O exchange): 6 1 .02 (d, J = 6.8 Hz, 3H), 1 .43 (s, 9H), 2.94 (m, 7H), 2.98 - 3.02 (m, 2H), 3.25 (s, 3H), 3.74 (m, 2H), 4.20 - 4.24 (m, 1 H), 4.45 (s, br, 1 H), 4.65 (d, J= 17.2 Hz, 1 H), 6.12 - 6.16 (m,1 H), 6.64 - 6.71 (m, 1 H), 7.12 (d, J= 8.0 Hz, 1 H), 7.19 (s, 1 H), 7.23 (d, J= 8.0 Hz, 1 H).

[0539] LCMS (Method N): 1.763 min, MS ES+: 388.4 (M+1).Step 8: (E)-4-(Dimethylamino)- / V-methyl- / V-(3-methyl-1 ,2,3,4-tetrahydroisoquinolin-7-yl) but-2-enamide hydrochloride

[0540] The title compound was prepared from fe / Y-butyl-(E)-7-(4-(dimethylamino)- / V-methylbut-2- enamido)-3-methyl-3,4-dihydroisoquinoline-2(1 H)-carboxylate (Step 7) and 4M HCI in dioxane analogously to Example 6, Step 2.

[0541] 1H NMR (DMSO-d6, 400 MHz, D2O exchange): 1 .41 (d, J = 8 Hz, 3H), 2.65 - 2.66 (m, 6H), 2.78- 2.90 (m, 1 H), 3.04 - 3.09 (m, 1 H), 3.25 (s, 3H), 3.53 - 3.67 (m, 2H), 4.29 (s, 2H), 6.12 - 6.15 (m, 1 H), 6.65- 6.72 (m, 1 H), 7.20 - 7.29 (m, 3H).

[0542] LCMS (Method N): 0.326 min, MS ES+: 288.22 (M+1).Intermediate K: (3-Chloro-6-hvdroxy-4-methoxy-2-methylphenyl)(4-(methylamino)isoindolin-2- vDmethanoneStep 1 : N-Methylisoindolin-4-amine hydrochloride

[0543] A stirred solution of fe / Y-butyl-4-(methylamino)isoindoline-2-carboxylate (Intermediate H ) (5.0 g, 20.1 mmol, 1 eq.) in DCM (50 mL) was treated dropwise with 4M HCI in dioxane (25 mL) at 0°C and allowed to warm to room temperature, stirring for 2 h. The resulting mixture was concentrated under reduced pressure and trituration with n-pentane (3 x 10 mL) afforded the title compound as an off-white solid (5.0 g, Yield: Quantitative).

[0544] 1H NMR (DMSO-d6, 400 MHz): 6 2.75 (s, 3H), 4.37 - 4.44 (m, 4H), 6.64 (d, J= 7.6 Hz, 1 H), 6.74 (d, J= 7.2 Hz, 1 H), 7.23 (t, J= 8.0 Hz, 1 H), 9.94 (bs, 2H, D2O exchangeable).

[0545] LCMS (Method A): 0.667 min, MS: ES+ 148.8 (M+1).Step 2: (3-Chloro-6-hydroxy-4-methoxy-2-methylphenyl) (4-(methylamino) isoindolin-2-yl) methanone

[0546] Performed in 2 parallel batches, each of 1 .5 g scale: To a stirred solution of 3-chloro-6-hydroxy- 4-methoxy-2-methylbenzoic acid (Intermediate D) (1.5 g, 8.15 mmol, 1.0 eq.) in DMF (15mL) were added EDC.HCI (1.989 g, 12.22 mmol, 1.5 eq.) and HOAt (1.427 g, 12.22 mmol, 1.5 eq.) and the mixture was stirred at room temperature for 15 mins. / V-Methylisoindolin-4-amine hydrochloride (Step 1) (1 .277 g, 8.15 mmol, 1 .0 eq.) and NMM (4.6 mL, 48.91 mmol, 6.0 eq.) were added and stirring continued at room temperature for 1 h. The resulting mixture was poured into ice cold water (60 mL) and extracted with EtOAc (3 x 40 mL). The combined organic extracts were dried over Na2SO4, filtered and concentrated under reduced pressure. The crude material was purified by chromatography on silica (compound eluted at 80% EtOAc in hexane) to yield (3-chloro-6-hydroxy-4-methoxy-2-methylphenyl) (4-(methylamino) isoindolin-2- yl) methanone as a white solid (1.2 g, Yield: 4.81 %,).

[0547] 1H NMR (DMSO-d6, 400 MHz): 6 2.17 (s, 3H), 2.61 - 2.73 (m, 3H), 3.81 (s, 3H), 4.17 - 4.48 (m, 2H), 4.72 (s, 2H), 5.31-5.52 (m, 1 H, D2O exchangeable), 6.35 - 6.45 (m, 1 H), 6.53 - 6.59 (m, 2H), 7.06 - 7.14 (m, 1 H), 10.01 (s, 1 H).

[0548] LCMS (Method N): 2.092 min, MS: ES+ 347.4,349.4 (M, M+2).

[0549] HPLC (Method H): 7.34 min.Intermediate KA: (4-Aminoisoindolin-2-yl)(3-chloro-6-hvdroxy-4-methoxy-2-methylphenyl)methanoneStep 1 : lsoindolin-4-amine hydrochloride

[0550] The title compound was prepared from te / Y-butyl 4-aminoisoindoline-2-carboxylate (CAS 871013- 98-8) and 4M HCI in dioxane analogously to Intermediate K, Step 1 .

[0551] 1H NMR (DMSO-d6, 400 MHz): 6 4.48 - 4.55 (m, 4H), 7.06 - 7.08 (m, 2H), 7.28 - 7.32 (m, 1 H),10.05 (bs, 1 H).

[0552] LCMS (Method: N): 2.44 min, MS ES+: 135.01 (M+1).Step 2: (4-Aminoisoindolin-2-yl)(3-chloro-6-hydroxy-4-methoxy-2-methylphenyl)methanone

[0553] The title compound was prepared from 3-chloro-6-hydroxy-4-methoxy-2-methylbenzoic acid (Intermediate D) and isoindolin-4-amine hydrochloride (Step 1) analogously to Intermediate K, Step 2.

[0554] 1H NMR (DMSO-d6, 400 MHz, D2O exchange): 6 2.17 (s, 3H), 3.81 (s, 3H), 4.19 - 4.69 (m, 4H), 6.38 - 6.54 (m, 3H), 6.92 - 7.00 (m, 1 H).

[0555] LCMS (Method: N): 1.391 min, MS ES+: 333.0, 335.0 (M, M+2).Intermediate L: (E)-4-((f-Butoxycarbonyl)amino)but-2-enoic acidStep 1 : (E)-4-Aminobut-2-enoic acid

[0556] To (E)-4-bromobut-2-enoic acid (CAS 13991-36-1 , 1.0 g, 6.06 mmol, 1.0 eq.) was added aqueous ammonia solution (10 mL) and the reaction mixture was stirred at room temperature for 16 h. The resulting mixture was concentrated under reduced pressure and azeotroping with n-hexane to yield (E)-4- aminobut-2-enoic acid as a white solid (0.85 g, Yield: quantitative). The material was used in the next step without further purification.

[0557] 1H NMR (DMSO-d6, 400 MHz, D2O exchange): 6 3.27 (d, J= 5.6 Hz, 1 H), 3.51 (d, J= 5.6 Hz, 1 H), 5.85 - 5.97 (m, 1 H), 6.63 - 6.93 (m, 1 H).

[0558] LCMS (Method N): 0.243 min, MS: ES+ 102.12 (M+1).Step 2: (E)-4-((fe / Y-Butoxycarbonyl) amino) but-2-enoic acid

[0559] To a stirred solution of ((E)-4-aminobut-2-enoic acid (Step 1) (0.85 g, 8.40 mmol, 1.0 eq.) in 1 :1 THF: water (42.50 mL) were added sodium carbonate (1 .78 g, 16.81 mmol, 2 eq.) and Boc anhydride (3.66 g, 16.81 mmol, 2 eq) and the reaction mixture was stirred at room temperature for 15 h. The resulting mixture was acidified to pH 2 with 1 N HCI and extracted with EtOAc (3 x 50 mL). The combined organic extracts were dried over Na2SO4, filtered and concentrated under reduced pressure. The crude material was purified by chromatography on silica (product eluted in 6% MeOH: MDC) yielding (E)-4-((fe / Y- butoxycarbonyl) amino) but-2-enoic acid as an off-white solid (0.30 g, Yield: 19%).

[0560] 1H NMR (DMSO-d6, 400 MHz, D2O exchange): 6 1.39 (s, 9H), 3.71 (s, 2H), 5.75 (d, J= 15.6 Hz, 1 H), 6.70 - 6.76 (m, 1 H), 7.18 (s, 1 H).Intermediate M: te / Y-Butyl-(E)-2-(3-ethoxy-3-oxoprop-1 -en-1-yl)pyrrolidine-1 -carboxylateO°C-rt, 1h

[0561] To a cooled (0°C) solution ethyl 2-(diethoxyphosphoryl) acetate (CAS: 867-13-0) (12.37 g, 55.22 mmol, 1 eq.) in THF (100 mL) was added potassium f-butoxide (5.61 g, 50.09 mmol, 1.0 eq.) and the mixture stirred at 0°C for 30 mins. te / Y-Butyl 2-formylpyrrolidine-1-carboxylate (CAS: 117625-90-8) (10 g, 50.12 mmol, 1 eq.) was added at 0°C and the reaction mixture was allowed to warm to room temperature stirring for 1 h. The resulting mixture was poured into ice cold water (500 mL) and extracted with EtOAc (3 x 300 mL). The combined organic extracts were dried over Na2SC>4, filtered, and concentrated under reduced pressure. The crude material was purified by chromatography on silica (product eluted at 25% EtOAc: hexane) yielding te / Y-butyl (E)-2-(3-ethoxy-3-oxoprop-1-en-1-yl)pyrrolidine-1 -carboxylate as a colourless oil (10 g, Yield: 74%).

[0562] 1H NMR (DMSO, 400 MHz): 6 1.21 (t, J= 7.2 Hz, 3H), 1.38 (d, J= 16.4 Hz, 9H), 1.74 - 1.78 (m, 3H), 2.04 - 2.07 (m, 1 H), 3.25 - 3.29 (m, 2H), 4.09 - 4.15 (q, br, 2H), 4.31 - 4.37 (m, 1 H), 5.74 - 5.78 (m, 1 H), 6.75 - 6.79 (m, 1 H).

[0563] LCMS (Method N): 1.730 min, MS: ES+ 170.1 (M+1).Intermediate N: 5-Cvclopropyl-2-hvdroxy-4-methoxybenzoic acidStep 1 : Methyl 5-cyclopropyl-2,4-dimethoxybenzoate90°C,32h

[0564] A stirred solution of methyl 5-bromo-2,4-dimethoxybenzoate (CAS: 39503-51-0) (4.0 g, 14.540 mmol, 1.0 eq.) and potassium cyclopropyl trifluoro borate (CAS: 1065010-87-8) (3.2g, 21.810 mmol, 1.5 eq.) in dioxane: H2O (40 mL) were placed in a sealed tube. K2CO3 (4.0 g, 29.080 mmol, 2.0 eq.) was added and the mixture was purged with N2 gas for 20 mins at room temperature. PdCl2(dppf) (0.532 g, 0.727 mmol, 0.05 eq.) was added and the reaction mixture was heated to 90°C for 32 h. The resulting mixture was filtered through Celite® and the filtrate was concentrated under reduced pressure. The crude material was purified by chromatography on silica (product eluted at 18% ethyl acetate in hexane) to afford methyl 5-cyclopropyl-2,4-dimethoxybenzoate as a white solid. (1.1 g, 4.65 mmol, Yield: 32 %).

[0565] 1H NMR (DMSO-d6, 400 MHz): 6 0.52 - 0.54 (m, 2H), 0.81 - 0.86 (m, 2H), 1.92 - 1.97 (m, 1 H), 3.71 (s, 3H), 3.81 (s, 3H), 3.91 (s, 3H), 6.66 (s, 1 H), 7.21 (s, 1 H).

[0566] LCMS (Method N): 1.619 min, MS: 237.14 (M+1).Step 2: Methyl 5-cyclopropyl-2-hydroxy-4-methoxybenzoate

[0567] To a cooled (0°C) solution of methyl 5-cyclopropyl-2,4-dimethoxybenzoate (Step 1) (1 .0 g, 4.232 mmol, 1.0 eq.) in DCM (15 mL) was added dropwise BCh (1 M in heptane) over the period of 10 mins and the reaction mixture was stirred at 0°C for 15 mins. The reaction was quenched with cold saturated NaHCCh solution (200 mL) and extracted with ethyl acetate (2 x 150 mL). The combined organic extracts were washed with saturated brine solution (1 x 100 mL), dried over Na2SC>4, filtered, and concentrated under reduced pressure. The crude material was purified by chromatography on silica (product eluted at 6% ethyl acetate in hexane) to afford the methyl 5-cyclopropyl-2-hydroxy-4-methoxybenzoate as a white solid. (0.67 g, Yield: 65%).

[0568] 1H NMR (DMSO-d6, 400 MHz): 6 0.49 - 0.53 (m, 2H), 0.82 - 0.85 (m, 2H), 1.89 - 1.93 (m, 1 H), 3.85 (d, J= 4.4 Hz, 6H), 6.54 (s, 1 H), 7.23 (s, 1 H), 10.69 (s, 1 H).Step 3: 5-Cyclopropyl-2-hydroxy-4-methoxybenzoic acid

[0569] To a stirred solution of methyl 5-cyclopropyl-2-hydroxy-4-methoxybenzoate (Step 2) (0.67 g, 3.014 mmol, 1.0 eq.) in MeOH: water (1 :1) (30 vol) was added LiOH.F (1 .27 g, 30.15 mmol, 10.0 eq.) and the reaction mixture was heated to 65°C for 6 h. The resulting mixture was acidified with sat. KHSC solution (pH 2-3) and extracted with ethyl acetate (2 x 150 mL). The combined organic extracts were further washed with saturated brine (1 x 100 mL) solution, dried over Na2SC>4, filtered, and concentrated under reduced pressure to afford 5-cyclopropyl-2-hydroxy-4-methoxybenzoic acid as a white solid (0.59 g, Yield: 93%).

[0570] 1H NMR (DMSO-d6, 400 MHz): 6 0.50 (d, J= 4.4 Hz, 2H), 0.80 - 0.84 (m, 2H), 1 .89 - 1.91 (m, 1 H), 3.84 (s, 3H), 6.47 (s, 1 H), 7.21 (s, 1 H), 12.60 (bs, 2H).

[0571] LCMS (Method N): 1.499 min, MS: 207.1 (M-1).Intermediate O: ((E)-4-(Dimethylamino)- / V-ethyl- / V-isoindolin-4-yl-but-2-enamide dihydrochlorideStep 1 : te / Y-Butyl 4-[[(E)-4-(dimethylamino)but-2-enoyl]-ethyl-amino]isoindoline-2-carboxylate

[0572] A solution of fe / Y-butyl-4-(ethylamino)isoindoline-2-carboxylate (CAS: 2002887-29-6, 554 mg, 2.11 mmol, 1.0 eq) in MeCN (15 mL) at 0°C was treated dropwise with a solution of (E)-4- (dimethylamino)but-2-enoyl chloride hydrochloride (CAS: 1055943-40-2, 428 mg, 2.3 mmol, 1.1 eq) in MeCN (5 mL) and the reaction mixture was stirred for 4 h. Additional MeCN (10 mL) was added to provide a clear solution and stirring continued for a further 10 mins. The resulting mixture was concentrated under reduced pressure. The crude residue was purified by chromatography on silica eluting with a gradient of 0 - 20% MeOH in DCM to afford the title compound (360 mg, Yield: 46%) which was used without further purification.Step 2: ((E)-4-(Dimethylamino)- / V-ethyl- / V-isoindolin-4-yl-but-2-enamide dihydrochloride

[0573] A solution of fe / Y-butyl-4-[[(E)-4-(dimethylamino)but-2-enoyl]-ethyl-amino]isoindoline-2- carboxylate (Step 1) (360 mg, 0.964 mmol) in MeOH (5 mL) was treated with 3M HCI in CPME (7.0 mL, 21.1 mmol, 10.0 eq) and stirred at room temperature for 2 h. The resulting mixture was concentrated underreduced pressure and azeotroped with toluene three times to afford a grey solid. The solid was dried under high vacuum overnight to afford the title compound (280 mg, Yield: 38%) as a grey solid which was used without further purification.

[0574] MS: ES+ 274.1 (M+1).

[0575] 1H NMR (400 MHz, DMSO) 6 1.07 - 1 .02 (m, 3H), 2.68 - 2.63 (m, 6H), 3.86 - 3.67 (m, 4H), 4.46 - 4.33 (m, 2H), 4.61 - 4.55 (m, 2H), 6.04 - 5.99 (m, 1 H), 6.79 - 6.70 (m, 1 H), 7.30 - 7.14 (m, 2H), 7.53 - 7.45 (m, 1 H), 10.03 - 10.00 (m, 1 H), 10.45 - 10.40 (m, 1 H), 10.73 (s, 1 H).Intermediate P: (E)-4-[Ethyl(methyl)amino1- / V-isoindolin-4-yl- / V-methyl-but-2-enamide dihydrochlorideStep 1 : Ethyl (E)-4-[ethyl(methyl)amino]but-2-enoate

[0576] To a suspension of potassium carbonate (1.0 g, 7.25 mmol, 2.0 eq) and / V-ethylmethylamine (0.56 mL, 6.53 mmol, 1 .80 eq) in THF (8.5 mL) was added a solution of ethyl 4-bromocrotonate (CAS: 37746-78-4, 0.50 mL, 3.63 mmol, 1 .0 eq) in THF (1 .0 mL) dropwise. The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water (20 mL) and extracted with ethyl acetate (2 x 30 mL). The combined organic extracts were washed with brine (30 mL), dried over MgSC , filtered and concentrated under reduced pressure to afford the title compound (557 mg, 3.25 mmol, 90%) as an orange oil.

[0577] MS: ES+ 172.1 (M+1).

[0578] 1H NMR (400 MHz, CDCb) 6 1 .07 (t, J=7.2 Hz, 3H), 1 .29 (t, J=7.2 Hz, 3H), 2.23 (s, 3H), 2.44 (q, J=7.1 Hz, 2H), 3.14 (dd, J=1 .6, 6.3 Hz, 2H), 4.19 (q, J=7.2 Hz, 2H), 5.97 (td, J=1 .6, 15.7 Hz, 1 H), 6.97 (td, J=6.3, 15.7 Hz, 1 H).Step 2: (E)-4-[Ethyl(methyl)amino]but-2-enoic acid

[0579] To a stirred solution of ethyl (E)-4-[ethyl(methyl)amino]but-2-enoate (Step 1) (557 mg, 3.25 mmol, 1.0 eq) in THF (13 mL) was added 2M sodium hydroxide (4.9 mL, 9.76 mmol, 3.0 eq) and the reaction mixture was stirred at room temperature for 16 h. The pH was adjusted to pH 6 with 2N HCI and the mixture was concentrated under reduced pressure. 10% methanol in DCM (60 ml) was added and the resulting suspension was stirred for 2 h, filtered and concentrated under reduced pressure to afford the title compound (483 mg, Yield: Quantitative) as a pale brown gum.

[0580] 1H NMR (400 MHz, CDCh) 6 1 .26 (t, J=7.3 Hz, 3H), 2.57 (s, 3H), 2.97 - 2.90 (m, 2H), 3.52 - 3.48 (m, 2H), 6.10 (d, J=15.6 Hz, 1 H), 6.82 - 6.73 (m, 1 H), 8.06 (s, 2H).Step 3: te / Y-Butyl 4-[[(E)-4-[ethyl(methyl)amino]but-2-enoyl]-methyl-amino]isoindoline-2-carboxylate

[0581] Oxalyl chloride (0.079 mL, 0.908 mmol, 1.3 eq) followed by 2 drops of DMF was added to a suspension of (E)-4-[ethyl(methyl)amino]but-2-enoic acid (Step 2) (100 mg, 0.70 mmol, 1.0 eq) in acetonitrile (6.5 mL) at 0°C. After 10 minutes, the solution was allowed to warm to ambient temperature and stirred for 2.5 h. This solution was then added to a solution of fe / Y-butyl-4-(methylamino)isoindoline-2- carboxylate (193 mg, 0.698 mmol, 1 .0 eq) (Intermediate H) in MeCN (6.5 mL) at 0°C. The reaction mixture was allowed to warm to room temperature whilst stirring for 18 h. The resulting mixture was concentrated under reduced pressure and the crude residue was purified by chromatography on silica, eluting with a gradient of 0 to 15% (0.1 % 7N NH3 / MeOH) in DCM afford the title compound (65 mg, Yield: 25%).

[0582] MS: ES+ 374.2 (M+1).Step 4: (E)-4-[Ethyl(methyl)amino]- / V-isoindolin-4-yl- / V-methyl-but-2-enamide dihydrochloride

[0583] A stirred solution of te / Y-butyl 4-[[(E)-4-[ethyl(methyl)amino]but-2-enoyl]-methyl- amino]isoindoline-2-carboxylate (Step 3) (70 mg, 0.187 mmol, 1 .0 eq) in DCM (1.2 mL) was treated dropwise with 3M HCI in CPME (0.62 mL, 1.87 mmol, 10.0 eq). The reaction mixture was stirred at room temperature for 3 h. Additional 3M HCI in CPME (0.62 mL, 1.87 mmol, 10.0 eq) was added and stirring continued for another hour. The resulting mixture was concentrated under reduced pressure and dried under high vacuum to afford the title compound (73 mg, Yield: Quant.) as a dark green solid.

[0584] MS: ES+ 274.1 (M+1).Intermediate Q: (E)-4-(Ethyl(methyl)amino)- / V-(isoindolin-4-yl)but-2-enamide dihydrochloride

[0585] The title compound was prepared from f-butyl 4-aminoisoindoline-2-carboxylate (CAS: 871013- 98-8) and (E)-4-[ethyl(methyl)amino]but-2-enoic acid analogously to Intermediate P, Steps 3 and 4.

[0586] 1H NMR (400 MHz, DMSO) 6 1.25 (t, J=7.3 Hz, 3H), 2.74 - 2.70 (m, 3H), 3.19 - 3.02 (m, 2H), 4.01 - 3.85 (m, 2H), 4.55 - 4.46 (m, 4H), 6.60 (d, J=15.4 Hz, 1 H), 6.90 - 6.81 (m, 1 H), 7.22 (d, J=7.6 Hz, 1 H), 7.37 (t, J=7.8 Hz, 1 H), 7.52 (d, J=7.9 Hz, 1 H), 9.87 - 9.79 (m, 2H), 10.43 (s, 1 H), 10.67 - 10.67 (m, 1 H).

[0587] MS: ES+ 260.1 (M+1).Intermediate R: 3-Bromo-6-hvdroxy-4-methoxy-2-methyl-benzoic acidStep 1 : 6-Bromo-7-methoxy-2,2,5-trimethyl-1 ,3-benzodioxin-4-one

[0588] A solution of 7-methoxy-2,2,5-trimethyl-1 ,3-benzodioxin-4-one (393 mg, 1.77 mmol, 1.0 eq) (CAS: 1628535-38-5) and / V-bromosuccinimide (472 mg, 2.65 mmol, 1.50 eq) in DCM (10 mL) and MeOH (5 mL) was stirred at ambient temperature overnight. The resulting mixture was concentrated under reduced pressure and the crude residue was purified by chromatography on silica eluting with 0 - 30% EtOAc in cyclohexane to afford the title compound (425 mg, Yield: 80%) as an off-white solid.

[0589] MS: ES+ 300.9, 303.0 (M+1).

[0590] 1H NMR (400 MHz, CDCb) 6 1.71 (s, 6H), 2.84 (s, 3H), 3.93 (s, 3H), 6.38 (s, 1 H).Step 2: 3-Bromo-6-hydroxy-4-methoxy-2-methyl-benzoic acid

[0591] A solution of 6-bromo-7-methoxy-2,2,5-trimethyl-1 ,3-benzodioxin-4-one (Step 1) (540 mg, 1.79 mmol, 1.0 eq) in THF (10 mL) was treated with 1 M lithium hydroxide (5.4 mL, 5.38 mmol, 3.0 eq) and stirred at room temperature for 18 h. The resulting mixture was diluted with water (20 mL) and washed with EtOAc (20 mL). The aqueous phase was acidified to pH 1-2 using 1 M HCI and extracted with EtOAc (3 x 30 mL). The combined organic extracts were washed with brine, dried over MgSO4 and concentrated under reduced pressure to afford the title compound (408 mg, Yield: 87%) as an off-white solid.

[0592] MS: ES+ 259.0, 261.0 (M+1).

[0593] 1H NMR (400 MHz, DMSO) 6 2.41 (s, 3H), 3.81 (s, 3H), 6.49 (s, 1 H).Intermediate S: (E)-4-(Dimethylamino)- / V-(isoindolin-4-yl)- / V-methylbut-2-enamide dihydrochloride

[0594] A solution of fe / Y-butyl-4-(methylamino)isoindoline-2-carboxylate (Intermediate H) (1.2 g, 4.7 mmol, 1 .0 eq) in acetonitrile (40 mL) was added dropwise to a solution of (E)-4-(dimethylamino)but-2-enoyl chloride (CAS 1056149-69-9, 860 mg, 4.7 mmol, 1 .0 eq) in acetonitrile (10 mL) at 0°C in an ice / water bath. The mixture was allowed to warm to room temperature stirring for 16 h. The resulting mixture was concentrated under reduced pressure and the crude residue was suspended in 4M HCI in dioxane (40 mL). The mixture was stirred for 2-3 h then concentrated under reduced pressure. The crude residue was triturated with / PrOH then isolated by filtration, washed with further / PrOH then dried under high vacuum. The mother liquors from the trituration were concentrated under reduced pressure, dissolved in / PrOH (10 mL) then added dropwise to Et2O (100 mL) with rapid stirring. The precipitated solid was isolated by filtration, dried under high vacuum then both batches of solid were combined to afford the title compound (1.3 g, Yield: 87%).

[0595] MS: ES+ 260.2 (M+1).Intermediate T: (E)-4-(Dimethylamino)- / V-(isoindolin-4-yl)-but-2-enamide dihydrochloride

[0596] The title compound was prepared from (E)-4-(dimethylamino)but-2-enoyl chloride and fe / Y-butyl 4-aminoisoindoline-2-carboxylate (CAS: 871013-98-8) analogously to Intermediate S.

[0597] MS: ES+ 246 (M+1).Intermediate U: 3-Chloro-2-fluoro-6-hvdroxy-4-methoxybenzoic acidStep 1 : 3-Chloro-2-fluoro-4,6-dimethoxybenzaldehyde

[0598] To a stirred solution of 2-fluoro-4,6-dimethoxybenzaldehyde (prepared from 1-fluoro-3,5- dimethoxybenzene (CAS: 52189-63-6) and POCh analogously to Intermediate C Step 1) (75.0 g, 407.23 mmol, 1 eq.) in DMF (750 mL) was added NCS (54.37 g, 407.23 mmol, 1 .0 eq.); the reaction mixture was stirred at room temperature for 16 h. then filtered and dried under vacuum. The crude material wastriturated with n-hexane and diethyl ether (5 x 100 mL) to yield 3-chloro-2-fluoro-4,6- dimethoxybenzaldehyde as a white solid (55g, Yield: 56.3 %).

[0599] 1H NMR (DMSO-d6, 400 MHz): 6 4.00 (s, 3H), 4.05 (s, 3H), 6.78 (s, 1 H), 10.16 (s, 1 H).Step 2: 3-Chloro-2-fluoro-4,6-dimethoxybenzoic acid

[0600] Performed in 2 parallel batches, each of 10 g scale: To a stirred solution of 3-chloro-2-fluoro-4,6- dimethoxybenzaldehyde (Step 1) (10 g, 45.7 mmol, 1 eq.) in DMSO (50 mL) were slowly added a saturated solution of NaH2PO4 (38.42 g, 320.20 mmol, 7.0 eq) and NaCIC>2 (28.95 g, 320.20 mmol, 7.0 eq). The reaction mixture was stirred at room temperature for 16 h. The resulting mixture was diluted with water (500 mL) and extracted with ethyl acetate (4 x 500 mL). The combined organic extracts were dried over Na2SC>4, filtered, and concentrated under vacuum yielding 3-chloro-2-fluoro-4,6-dimethoxybenzoic acid as a white solid (15 g, yield: 51 .2%). The crude material was used without further purification.

[0601] 1H NMR (DMSO-d6, 400 MHz): 6 3.87 (s, 3H), 3.96 (s, 3H), 6.71 (s, 1 H), 13.33 (s, 1 H).Step 3: 3-Chloro-2-fluoro-6-hydroxy-4-methoxybenzoic acid

[0602] To a cooled (0°C) solution of 3-chloro-2-fluoro-4,6-dimethoxybenzoic acid (Step 2) (2 g, 8.50 mmol, 1 eq.) in DCM (10 mL) was added dropwise BBrs (1 M in DCM) (10 mL). The reaction mixture was allowed to warm to room temperature and stirred for 16 h. The resulting mixture was concentrated under vacuum and the crude residue was diluted with ice-cold water (50 mL) causing precipitation of a solid. The solid was collected by filtration and dried under high vacuum to yield 3-chloro-2-fluoro-6-hydroxy-4- methoxybenzoic acid as an off-white solid (1 .2 g, Yield: 63.0%).

[0603] 1H NMR (DMSO-d6, 400 MHz): 6 3.91 (s, 3H), 6.59 (d, J= 1.6 Hz, 1 H), 12.18 (bs, 2H).

[0604] LCMS (Method A): 1.800 min, MS: ES- 219 (M-1).Biological assaysFluorescence polarisation assay for PMS2

[0605] Test compounds, as 10 mM DMSO stocks, were dispensed into a Black Fluotrac 200 384 well medium binding plate (Greiner Bio-One, item number 781076) using a Labcyte Echo acoustic liquid handler. For single point screening, test compounds were added to wells in columns 1-22 whilst DMSO was added to wells in columns 23 and 24 in order to normalise the plate. For potency determination, serial dilutions of test compounds were added to wells in columns 3-22 and DMSO volume was normalised across the plate.

[0606] 20 pL of a 2 x solution (20 nM) of recombinant / V-terminal PMS2 (residues 1-365) in assay buffer (25 mM HEPES, pH 7.5, 250 mM NaCI, 10 mM MgCI2, 0.01 % Triton X-100, 5 mM Dithiothreitol) was added to all wells in columns 2-23 for potency determination or columns 1-23 for single point screening. 20 pL assay buffer was added to all wells in columns 1 and 24 (column 24 only for single point screening) using a MultiDrop Combi (ThermoFisher). Plates were centrifuged for 1 minute at 250 xg and were incubated at room temperature for 30 minutes prior to the addition of 20 pL of 2 x (20 nM) of 5-((5-(4-((2-(2,4-dihydroxy- 5-isopropylbenzoyl)isoindolin5-yl)methyl)piperazin-1-yl)pentyl)carbamoyl)-2-(6-(dimethylamino)-3- (dimethyliminio)-3 / 7-xanthen-9-yl)benzoate (referred to hereinafter as “probe compound”) in assay buffer (prepared from a 100 pM DMSO stock) with a MultiDrop Combi (ThermoFisher). The final concentration of / V-terminal PMS2 was 10 nM and the final concentration of probe compound was 5 nM.

[0607] Compound plates were centrifuged for 1 minute at 250 xg for 1 minute and were incubated at room temperature for 1 hour before being read on a PheraStar FSX (fitted with 384-well aperture spoon and 540 590 590 FP optic module). The gain and focus were adjusted before each plate was read so that the polarisation of a no enzyme control (column 24) was equal to 35 mP. Data were normalised against the no inhibitor controls (column 23) and no enzyme controls (column 24).Data obtained in this assay is shown in Table A1 shown belowTable A1* >1 pM** 0.1 - 1 pM*** less than 0.1 pMREFERENCES1 . Martin-Lopez, J.V. and R. Fishel, The mechanism of mismatch repair and the functional analysis of mismatch repair defects in Lynch syndrome. Fam Cancer, 2013. 12(2): p. 159-68.2. Liu, D., G. Keijzers, and L.J. Rasmussen, DNA mismatch repair and its many roles in eukaryotic cells. Mutat Res, 2017. 773: p. 174-187.3. Lynch, H.T., et al., Review of the Lynch syndrome: history, molecular genetics, screening, differential diagnosis, and medicolegal ramifications. Clin Genet, 2009. 76(1): p. 1-18.4. Shlien, A., et al., Combined hereditary and somatic mutations of replication error repair genes result in rapid onset of ultra-hypermutated cancers. Nat Genet, 2015. 47(3): p. 257-625. Sehgal, R., et al., Lynch syndrome: an updated review. Genes (Basel), 2014. 5(3): p. 497-5076. Willis, J.A., et al., Immune Activation in Mismatch Repair-Deficient Carcinogenesis: More Than Just Mutational Rate. Clin Cancer Res, 2019.7. Gubin, M.M. and R.D. Schreiber, CANCER. The odds of immunotherapy success. Science, 2015. 350(6257): p. 158-9.8. Kloor, M. and M. von Knebel Doeberitz, The Immune Biology of Microsatellite-Unstable Cancer. Trends Cancer, 2016. 2(3): p. 121-133.9. Giannakis, M., et al., Genomic Correlates of Immune-Cell Infiltrates in Colorectal Carcinoma. Cell Rep, 2016. 17(4): p. 1206.10. Lemery, S., P. Keegan, and R. Pazdur, First FDA Approval Agnostic of Cancer Site - When a Biomarker Defines the Indication. N Engl J Med, 2017. 377(15): p. 1409-1412.11. Le, D.T., et al., PD-1 Blockade in Tumors with Mismatch-Repair Deficiency. N Engl J Med, 2015. 372(26): p. 2509-20.12. Rizvi, N.A., et al., Cancer immunology. Mutational landscape determines sensitivity to PD-1 blockade in non-small cell lung cancer. Science, 2015. 348(6230): p. 124-8.13. Van Allen, E.M., et al., Genomic correlates of response to CTLA-4 blockade in metastatic melanoma. Science, 2015. 350(6257): p. 207-21 1.14. Rosenberg, J.E., et al., Atezolizumab in patients with locally advanced and metastatic urothelial carcinoma who have progressed following treatment with platinum-based chemotherapy: a singlearm, multicentre, phase 2 trial. Lancet, 2016. 387(10031): p. 1909-20.15. Hellmann, M.D., et al., Genomic Features of Response to Combination Immunotherapy in Patients with Advanced Non-Small-Cell Lung Cancer. Cancer Cell, 2018. 33(5): p. 843-852 e4.16. Rizvi, H., et al., Molecular Determinants of Response to Anti-Programmed Cell Death (PD)-1 and AntiProgrammed Death-Ligand 1 (PD-L1) Blockade in Patients With Non-Small-Cell Lung Cancer Profiled With Targeted Next-Generation Sequencing. J Clin Oncol, 2018. 36(7): p. 633-641.17. Carbone, D.P., et al., First-Line Nivolumab in Stage IV or Recurrent Non-Small-Cell Lung Cancer. N Engl J Med, 2017. 376(25): p. 2415-2426.18. Hellmann, M.D., et al., Nivolumab plus Ipilimumab in Lung Cancer with a High Tumor Mutational Burden. N Engl J Med, 2018. 378(22): p. 2093-2104.19. Hellmann, M.D., et al., Tumor Mutational Burden and Efficacy of Nivolumab Monotherapy and in Combination with Ipilimumab in Small-Cell Lung Cancer. Cancer Cell, 2018. 33(5): p. 853-861 e4.Germano, G., et al., Inactivation of DNA repair triggers neoantigen generation and impairs tumour growth. Nature, 2017. 552(7683): p. 116-120. Mandal, R., et al., Genetic diversity of tumors with mismatch repair deficiency influences anti-PD-1 immunotherapy response. Science, 2019. 364(6439): p. 485-491. Guan J., et al., MLH1 deficiency-triggered DNA hyperexcision by exonuclease 1 activates the cGAS- STI NG pathway. Cancer Cell. 2021 , 39 (1), 109 - 121 , Lu, C., et al. DNA sensing in mismatch repair-deficient tumor cells is essential for anti-tumor immunity. Cancer Cell. 2021 , 39 (1), 96 - 108. Manley, K. , et al., Msh2 deficiency prevents in vivo somatic instability of the CAG repeat in Huntington disease transgenic mice. Nat Genet, 1999. 23(4): p. 471-3.

Claims

CLAIMSA compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound is selected from any one of the following:

2. The compound according to claim 1 , or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound is selected from any one of the following:

3. The compound according to claim 1 or claim 2, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound is selected from any one of the following:

4. The compound according to any one of the preceding claims, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound is selected from any one of the following:

5. The compound according to any one of the preceding claims, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound is selected from any one of the following:

6. The compound according to any one of the preceding claims, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound is:

7. A pharmaceutical composition comprising a compound according to any one of the preceding claims, or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable excipient or carrier.

8. A compound according to any one of claims 1 -6, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition according to claim 7, for use in therapy.

9. A compound according to any one of claims 1 -6, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition according to claim 7, for use in the treatment of a proliferative disorder.

10. A compound according to any one of claims 1-6, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition according to claim 7, for use in the treatment of cancer.

11. A compound according to any one of claims 1-6, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition according to claim 7, for use in the treatment of a triplet repeat disorder.

12. The compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or the pharmaceutical composition for use of claim 11 , wherein the triplet repeat disorder is selected from the group consisting of Huntington’s disease (HD), myotonic dystrophy type 1 (DM1), fragile X syndrome type A (FRAXA), Friedreich’s ataxia (FRDA), and spinocerebellar ataxias (SCAs).

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