RU2830439C1 - Pharmaceutical combination based on prmt5 inhibitors - Google Patents

Abstract

FIELD: medicine; pharmaceutics.

SUBSTANCE: invention refers to pharmaceutics, namely to a pharmaceutical composition for treating cancer. Pharmaceutical combination for treating cancer in a subject is selected from: (a) compounds (1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 24) or a pharmaceutically acceptable salt thereof and afatinib; (b) compounds (1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 24) or a pharmaceutically acceptable salt thereof and BI-3406; (c) compounds (1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 24) or a pharmaceutically acceptable salt thereof and ulixertinib.

EFFECT: above described combinations have a synergistic effect in treating cancer in a subject.

2 cl, 10 dwg, 5 ex

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

This PCT application claims priority from Indian Provisional Patent Application No. 201921042899, filed on October 22, 2019, the contents of which are hereby incorporated by reference in their entirety.

All references, including without limitation patents, patent applications, non-patent references and products cited in this PCT application and their disclosures are hereby incorporated by reference in their entirety.

FIELD OF INVENTION

The present invention relates to a pharmaceutical combination for the treatment of cancer. In particular, the present invention relates to a pharmaceutical combination comprising a combination of an arginine N-methyl-transferase 5 (PRMT5) protein inhibitor of formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist or an immune checkpoint inhibitor/modulator, wherein the PRMT5 inhibitor is a compound of formula (I),

its tautomeric form, its stereoisomer, its pharmaceutically acceptable salt, their polymorphic forms or their solvates for use in the treatment and/or prevention of cancer.

LEVEL OF TECHNOLOGY

Protein methylation is a common post-translational modification that affects protein activity and its interactions with other biological molecules. N-methylation typically occurs at the nitrogen atoms of arginine, lysine, and histidine residues, and there are different families of enzymes that catalyze the methylation reaction, each specific to the amino acid residue to be methylated.

A family of 9 enzymes called protein arginine N-methyltransferases (PRMTs) are responsible for methylation of the guanidinium group of arginine. The guanidinium group of arginine contains two (2) terminal nitrogen atoms that undergo monomethylation or dimethylation. Depending on the type of dimethylation, the enzymes are further classified as type I or type II. Type I PRMTs catalyze monomethylation or asymmetric dimethylation, while type II enzymes catalyze symmetric dimethylation. Some of the substrates that undergo methylation are histones, Sm ribonucleoproteins, MRE11, and p53-binding protein 1.

Methylation of arginine side chains plays an important role in various cellular functions, including transcriptional activation as well as transcriptional repression, mRNA translation, pre-mRNA splicing, protein transport, and signal transduction. It also occurs in numerous substrates. Therefore, PRMT enzymatic activity influences cellular processes such as cell proliferation, DNA damage repair, as well as cell cycle and cell death. PRMT enzyme-mediated hypermethylation has been shown to lead to several disease states such as cancer (Nature Reviews Cancer, 2013, 13, p37; Cellular and Molecular Life Sciences, 2015, 72, p2041; Trends in Biochemical Sciences, 2011, 36, p633).

Currently, the most studied type II enzyme is PRMT5, which is conserved among eukaryotic organisms. PRMT5 overexpression is associated with carcinogenesis and decreased patient survival in several types of human malignancies (Cell Mol Life Sci., 2015, 72, p2041). PRMT5 directly interacts with proteins that are often deregulated or mutated in cancers and, therefore, is a putative oncogene (Mol Cell Biol, 2008, 28, p6262). PRMT5-mediated transcriptional suppression of tumor suppressor genes such as p53, RB-1, ST7, or upregulation of cyclin D1, CDK4, CDK6, eLF4E, MITF, FGFR3 are associated with oncogenesis in both solid tumors and hematological malignancies. PRMT5 is located in the nucleus as well as the cytoplasm, and its overexpression has been associated with a wide range of cancers, including but not limited to glioblastoma multiforme (Oncogene, 2017, 36, p263), prostate cancer (Oncogene, 2017, 36, p1223), and pancreatic cancer (Science, 2016, 351, p1214), mantle cell lymphoma (Nature Chemical Biology, 2015, 11, p432), non-Hodgkin lymphomas and diffuse large B-cell lymphoma (Journal of Biological Chemistry, 2013, 288, p35534), acute myeloid leukemia (Leukemia, 2018, 32, p499), acute lymphoblastic leukemia (AACR; Cancer Research 2017; 77 (13 Suppl): Abstract nr 1128), multiple myeloma (Leukemia, 2018, 32, p996), non-small cell lung cancer (The Biochemical Journal, 2012, 446, p235), small cell lung cancer (AACR; Cancer Research 2017; 77(13 Suppl): Abstract nr DDT02-04), breast cancer (Cell Reports, 2017, 21, p3498), triple-negative breast cancer (AACR; Cancer Res 2015; 75(15 Suppl): Abstract nr 4786), gastric cancer (International Journal of Oncology, 2016, 49, p1195), colorectal cancer (Oncotarget, 2015, 6, p22799), ovarian cancer (J Histochem Cytochem 2013, 61, p206), bladder cancer (Clinical Cancer Research, 2018, CCR-18-1270), hepatocellular carcinoma (Oncology Reports, 2018, 40, p536), melanoma (PLoS One, 2013, 8, e74710; J Clin Invest. 2018, 128, p517), sarcoma (Oncology Letters, 2018, 16, p2161), oropharyngeal squamous cell carcinoma (Oncotarget, 2017, 8, p14847), chronic myelogenous leukemia (J Clin Invest, 2016, 126, p3961), epidermal squamous cell carcinoma (Carcinogenesis, 2017, 38, p827), nasopharyngeal carcinoma (Oncology Reports, 2016, 35, p1703), neuroblastoma (Molecular Oncology, 2015, 9, p617), endometrial carcinoma (Gynecol Oncol., 2016, 140, p145), cervical cancer (Pharmazie, 2018, 73, p269). These results led to additional studies showing that inhibition of PRMT5 reduces cell proliferation (Molecular and Cellular Biology 2008, 28, p6262, The Journal of Biological Chemistry 2013, 288, p35534).

Arginine methyltransferase inhibitors were first reported in 2004 by Cheng et al in the Journal of Biological Chemistry – Vol. 279 (23), p.23892. Since then, various other compounds and substances have been reported that exhibit higher selectivity for either type I or type II arginine methyltransferases. Other publications that disclose small molecules as inhibitors of PRMT5 include: International PCT Publication Nos. WO2011077133, WO2011079236, WO2014100695, WO2014100716, WO2014100719, WO2014100730, WO2014100734, WO2014128465, WO2014145214, WO2015200677, WO2015200680, WO2015198229, WO2016022605, WO2016034671, WO2016034673, WO2016034675, WO2016038550, WO2016135582, WO2016145150, WO2016178870, WO2018065365, WO2019032859 and ACS Medicinal Chemistry Letters 2015, 6, p.408, Cell reports 2018, 24(10), p2643-2657; Proceedings of the National Academy, 2019, 116(36), p17990-18000; Blood Cancer Journal 2019, 33(12), p2898-2911 and Biorganic & Medicinal Chermistry Letters, 2019, 29(11), p1264-1269. Other publications that describe modulators of cellular activity are WO2018115380; WO2019051084; Cell Elsevier, 2018,172(3), p578.

However, there is a need to identify an improved pharmaceutical composition or methods for treating or preventing cancer in a subject in need thereof.

ESSENCE OF THE INVENTION

The inventions described and claimed herein are characterized by many features and aspects, including, but not limited to, those indicated or described or cited in this Summary of the Invention. The inventions described and claimed herein are not intended to include all of the features presented, and these inventions are not limited to the features or embodiments indicated in this Summary of the Invention, which are included solely for illustration and not limitation, or by them.

Taking into account the above problems, according to one aspect disclosed herein, the present invention relates to a pharmaceutical combination comprising a combination of a PRMT5 inhibitor of formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist or an immune checkpoint inhibitor/modulator, wherein the PRMT5 inhibitor is is a compound of formula (I),

its tautomeric form, its stereoisomer, its pharmaceutically acceptable salt, their polymorphic forms or their solvates, for use in the treatment and/or prevention of cancer.

According to one aspect disclosed herein, there is provided a pharmaceutical combination for the treatment and/or prevention of cancer in a subject, wherein the pharmaceutical combination comprises a PRMT5 inhibitor of formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator, wherein the compound of formula (I) is

Where

L ₁ is selected from -CR ^a R ^b -, -NR ^a -, S and O;

Z represents CH or N;

R ^a and R ^b are independently selected in each instance from hydrogen, substituted or unsubstituted alkyl, and substituted or unsubstituted cycloalkyl;

ring A is selected from

R ^c and R ^d are selected from substituted or unsubstituted alkyl or, together with the carbon atoms to which they are attached, form a C ₃ -C ₆ cycloalkyl ring;

R is selected from ^–NR4R5 , hydrogen, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted heteroaryl, and substituted or unsubstituted cycloalkyl ^;

R ¹ and R ² together with the carbon atoms to which they are attached form a bond to form –C=C- or R ¹ and R ² together with the carbon atoms to which they are attached form a cyclopropane ring;

R ^2' and R ^2a , which may be the same or different, are independently selected from hydrogen and substituted or unsubstituted alkyl;

R³independently selected at each occurrence from halogen, cyano, nitro, substituted or unsubstituted alkyl, -OR⁶, -NR⁷R⁸, substituted or unsubstituted cycloalkyl, -C(O)OH, -C(O)O-alkyl, -C(O)R⁹, -C(O)NR⁷R⁸, –NR⁷C(O)R⁹, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl and substituted or unsubstituted heterocyclyl;

R ⁴ and R ⁵ are independently selected from hydrogen, substituted or unsubstituted alkyl, and substituted or unsubstituted cycloalkyl;

R⁶ selected from hydrogen, substituted or unsubstituted alkyl, and substituted or unsubstituted cycloalkyl;

R ⁷ and R ⁸ are independently selected from hydrogen, substituted or unsubstituted alkyl, and substituted or unsubstituted cycloalkyl;

R ⁹ is selected from substituted or unsubstituted alkyl and substituted or unsubstituted cycloalkyl;

R¹⁰ selected from hydrogen, halogen and substituted or unsubstituted alkyl;

"n" is an integer between 0 and 4 inclusive;

if the alkyl group is substituted, it is substituted with 1-4 substituents independently selected from oxo (=O), halogen, cyano, cycloalkyl, aryl, heteroaryl, heterocyclyl, -OR ^7a , -C(=O)OH, -C(=O)O(alkyl), -NR ^8a R ^8b , -NR ^8a C(=O)R ^9a and –C(=O)NR ^8a R ^8b ;

if the heteroaryl group is substituted, it is substituted with 1-4 substituents independently selected from halogen, nitro, cyano, alkyl, haloalkyl, perhaloalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR^7a, -NR^8aR^8b, -NR^7aC(=O)R^9a, –C(=O)R^9a, –C(=O)NR^8aR^8b, -SO₂-alkyl, -C(=O)OH and -C(=O)O-alkyl;

if the heterocyclic group is substituted, it is substituted at either a ring carbon atom or a ring heteroatom, and if substituted at a ring carbon atom, it is substituted by 1-4 substituents independently selected from oxo (=O), halogen, cyano, alkyl, cycloalkyl, perhaloalkyl, -OR^7a, –C(=O)NR^8aR^8b, -C(=O)OH, -C(=O)O-alkyl, -N(H)C(=O)(alkyl), -N(H)R^8aand -N(alkyl)₂; and if the heterocyclic group is substituted at a nitrogen atom in the ring, then it is substituted with substituents independently selected from alkyl, cycloalkyl, aryl, heteroaryl, -SO₂(alkyl), –C(=O)R^9aAnd -C(=O)O(alkyl); if the heterocyclic group is substituted at the sulfur atom in the ring, then it is substituted by 1 or 2 oxo groups (=O);

R ^7a is selected from hydrogen, alkyl, perhaloalkyl and cycloalkyl;

each of R ^8a and R ^8b is independently selected from hydrogen, alkyl and cycloalkyl; and

R ^9a is selected from alkyl and cycloalkyl.

According to another aspect disclosed herein, the PRMT5 inhibitor of formula (I) in the pharmaceutical combination is a compound selected from the group consisting of

(1S,2R,5R)-3-(2-(2-amino-3-bromoquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 1);

(1S,2R,5R)-3-(2-(2-amino-3-chloroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 2);

(1S,2R,5R)-3-(2-(2-amino-3-bromoquinolin-7-yl)ethyl)-5-(4-amino-5-fluoro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 3);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(((2-aminoquinolin-7-yl)thio)methyl)cyclopent-3-ene-1,2-diol (compound 4);

(1S,2R,5R)-3-(((2-amino-3-chloroquinolin-7-yl)thio)methyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 5);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(((2-aminoquinolin-7-yl)(methyl)amino)methyl)cyclopent-3-ene-1,2-diol (compound 6);

(1S,2R,5R)-3-(1-(2-amino-3-bromoquinolin-7-yl)propan-2-yl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compounds 7a and 7b);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(1-((2-(methylamino)quinolin-7-yl)oxy)ethyl)cyclopent-3-ene-1,2-diol (compounds 8a and 8b);

(1S,2R,5R)-3-(((2-amino-3-chloro-5-fluoroquinolin-7-yl)oxy)methyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylcyclopent-3-ene-1,2-diol (compound 9);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(2-(2-(methylamino)quinolin-7-yl)ethyl)cyclopent-3-ene-1,2-diol (compound 10);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(2-(3-methylimidazo[1,2-a]pyridin-7-yl)ethyl)cyclopent-3-ene-1,2-diol (compound 11);

(1S,2R,5R)-3-(((2-amino-3-chloro-5-fluoroquinolin-7-yl)oxy)methyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-methylcyclopent-3-ene-1,2-diol (compound 12);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 13);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-methyl-1H-pyrrolo[3,2-c]pyridin-1-yl)cyclopent-3-ene-1,2-diol hydrochloride (compound 14);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(1H-pyrrolo[3,2-c]pyridin-1-yl)cyclopent-3-ene-1,2-diol (compound 15);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 16);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(((2-aminoquinolin-7-yl)amino)methyl)cyclopent-3-ene-1,2-diol (compound 17);

(1S,2R,5R)-3-(((2-amino-3-chloro-5-fluoroquinolin-7-yl)oxy)methyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 18);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(((2-(methylamino)quinolin-7-yl)oxy)methyl)cyclopent-3-ene-1,2-diol (compound 19);

(1S,2R,5R)-3-(1-((2-amino-3-chloro-5-fluoroquinolin-7-yl)oxy)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 20a and 20b);

(1S,2R,5R)-3-(2-(2-amino-3-chloroquinolin-7-yl)ethyl)-5-(4-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 21);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(2-(2-(cyclobutylamino)quinolin-7-yl)ethyl)cyclopent-3-ene-1,2-diol (compound 22);

(1S,2R,5R)-3-(2-(2-amino-3-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 23);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 24);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-6-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 25);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-8-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 26);

(1S,2R,5R)-3-(2-(2-amino-3,3-dimethyl-3H-indol-6-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 27);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(2-(2'-aminospiro[cyclobutan-1,3'-indol]-6'-yl)ethyl)cyclopent-3-ene-1,2-diol (compound 28);

(1S,2R,5R)-3-(2-(2-amino-3,5-dichloroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 29);

(1S,2R,5R)-3-(2-(2-amino-3-chloroquinolin-7-yl)ethyl)-5-(2-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 30);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 31);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 32);

(1S,2R,5R)-3-(1-(2-amino-3-chloro-5-fluoroquinolin-7-yl)propan-2-yl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compounds 33a and 33b);

(1S,2R,5R)-3-(1-(2-amino-3-chloroquinolin-7-yl)propan-2-yl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compounds 34a and 34b);

(1S,2R,5R)-3-(1-(2-amino-3-chloro-5-fluoroquinolin-7-yl)propan-2-yl)-5-(4-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compounds 35a and 35b);

(1S,2R,5R)-3-(1-(2-amino-3-chloro-5-fluoroquinolin-7-yl)propan-2-yl)-2-methyl-5-(4-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compounds 36a and 36b);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylcyclopent-3-ene-1,2-diol (compound 37);

(1S,2R,5R)-3-(1-(2-amino-3-chloro-5-fluoroquinolin-7-yl)propan-2-yl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylcyclopent-3-ene-1,2-diol (compound 38);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-2-methyl-5-(4-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 39);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-ethylcyclopent-3-ene-1,2-diol (compound 40);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-ethyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 41);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 42);

(1S,2R,5R)-3-(2-(2-amino-3-bromo-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 43);

(1S,2R,5R)-3-(2-(2-amino-3-bromo-5-fluoroquinolin-7-yl)ethyl)-5-(4-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 44);

(1S,2R,5R)-3-(1-(2-amino-3-bromo-5-fluoroquinolin-7-yl)propan-2-yl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compounds 45a and 45b);

(1S,2R,5R)-3-(2-(2-amino-6-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 46);

(1S,2R,5R)-3-(2-(2-amino-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 47);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2, 3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 48);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2-(methylamino)quinoline-7 -yl)ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 49);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2-(isopropylamino)quinolin-7-yl)ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 50);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2-(cyclobutylamino)quinoline-7 -yl)ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 51);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2-((cyclopropylmethyl)amino)quinolin-7-yl)ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 52);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2-amino-8-fluoroquinoline- 7-yl)ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 53);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-methylquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 54);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-isopropylquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 55);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-(1,1-difluoroethyl)quinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 56);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-cyclopropylquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 57);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-methoxyquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 58);

2-amino-7-(2-((1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,3-dihydroxybicyclo[3.1.0]hexan-1-yl)ethyl)quinoline-3-carbonitrile (compound 59);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-fluoroquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d] pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 60);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-chloroquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d] pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 61);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-chloro-6-fluoroquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2, 3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 62);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-chloro-8-fluoroquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2, 3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 63);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-bromo-6-fluoroquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2, 3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 64);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(3-methylimidazo[1,2-a]pyridin-7-yl)ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 65);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3,3-dimethyl-3H-indol-6-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 66);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-4-(4-amino-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 67);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-chloro-6-fluoroquinolin-7-yl)ethyl)-4-(4-amino-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 68);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-4-(4-methyl-7H-pyrrolo[2, 3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 69);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-4-(7H-pyrrolo[2,3-d] pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 70);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2'-aminospiro[cyclobutan-1,3'-indol]-6'-yl)ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 71);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-bromo-5-fluoroquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2, 3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 72);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-bromoquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d] pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 73);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2-aminoquinolin-7-yl) ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 74);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2-aminoquinazolin-7-yl) ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 75);

(1S,2R,3S,4R,5S)-1-((S)-1-(2-amino-3-bromoquinolin-7-yl)propan-2-yl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compounds 76a and 76b);

(1S,2R,3S,4R,5S)-1-((S)-2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)-1-cyclopropylethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compounds 77a and 77b);

(1S,2R,3S,4R,5S)-1-(1-(2-amino-3-chloro-5-fluoroquinolin-7-yl)propan-2-yl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compounds 78a and 78b);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2-aminoquinolin-7-yl)propyl)bicyclo[3.1.0]hexane-2,3-diol (compounds 79a and 79b);

(1R,2R,3S,4R,5S)-1-(((2-amino-3-bromoquinolin-7-yl)oxy)methyl)-4-(4-amino-7H-pyrrolo[2,3-d ]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 80);

(1S,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(((2-aminoquinolin-7-yl)thio )methyl)bicyclo[3.1.0]hexane-2,3-diol (compound 81);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-(4-fluorophenyl)quinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (connection 82)

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-(pyridin-3-yl)quinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 83) and

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-(3-methylisoxazol-4-yl)quinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 84),

or a pharmaceutically acceptable salt, hydrate or stereoisomer thereof.

According to another aspect disclosed herein, the PRMT5 inhibitor is (1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 24) or a pharmaceutically acceptable salt thereof.

According to another aspect disclosed herein, at least one targeting agent/modulator of cellular activity is an inhibitor of EGFR or oncogenic variants thereof. According to a further aspect disclosed herein, the EGFR inhibitor is afatinib, osimertinib, erlotinib or gefitinib.

According to another aspect disclosed herein, at least one targeting agent/modulator of cellular activity is an inhibitor of an SOS1 inhibitor. According to a further aspect disclosed herein, the SOS1 inhibitor is BI-3406.

According to another aspect disclosed herein, at least one targeting agent/modulator of cellular activity is an inhibitor of an ERK inhibitor. According to a further aspect disclosed herein, the ERK inhibitor is ulixertinib, MK-8353, or ravoxertinib.

According to another aspect disclosed herein, at least one targeting agent/modulator of cellular activity is a PARP inhibitor. According to a further aspect disclosed herein, the PARP inhibitor is olaparib, niraparib, rucaparib, or talazoparib (BMN-673).

According to another aspect disclosed herein, at least one targeted agent/modulator of cellular activity is a CDK4/6 inhibitor. According to a further aspect disclosed herein, the CDK4/6 inhibitor is palbociclib, ribociclib, and abemaciclib.

According to another aspect disclosed herein, at least one targeting agent/modulator of cellular activity is a MALT1 inhibitor. According to a further aspect disclosed herein, the MALT1 inhibitor is MI-2, JNJ-67856633.

According to another aspect disclosed herein, at least one targeting agent/modulator of cellular activity is a BTK inhibitor. According to a further aspect disclosed herein, the BTK inhibitor is ibrutinib, zanubrutinib, and acalabrutinib.

According to another aspect disclosed herein, a PRMT5 inhibitor compound is administered simultaneously, in parallel, sequentially, one after the other, alternately or separately with at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist or an immune checkpoint inhibitor/modulator.

According to another aspect disclosed herein, there is provided a pharmaceutical combination for treating and/or preventing cancer in a subject, wherein the pharmaceutical combination comprises

(a) a compound which is (1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 24), or a pharmaceutically acceptable salt thereof and

(b) afatinib.

According to another aspect disclosed herein, there is provided a pharmaceutical combination for treating and/or preventing cancer in a subject, wherein the pharmaceutical combination comprises

(a) a compound which is (1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 24), or a pharmaceutically acceptable salt thereof and

(b) BI-3406.

According to another aspect disclosed herein, there is provided a pharmaceutical combination for treating and/or preventing cancer in a subject, wherein the pharmaceutical combination comprises

(a) a compound which is (1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 24), or a pharmaceutically acceptable salt thereof and

(b) ulixertinib.

According to another aspect disclosed herein, there is provided a method for treating and/or preventing cancer in a human subject in need thereof, the method comprising administering to the human subject a therapeutically effective amount of any of the pharmaceutical combinations disclosed herein.

According to another aspect disclosed herein, the cancer is glioblastoma multiforme, prostate cancer, pancreatic cancer, mantle cell lymphoma, non-Hodgkin's lymphomas and diffuse large B-cell lymphoma, acute myeloid leukemia, acute lymphoblastic leukemia, multiple myeloma, non-small cell lung cancer, small cell lung cancer, breast cancer, triple-negative breast cancer, gastric cancer, colorectal cancer, ovarian cancer, bladder cancer, hepatocellular carcinoma, melanoma, sarcoma, oropharyngeal squamous cell carcinoma, chronic myelogenous leukemia, epidermal squamous cell carcinoma, nasopharyngeal carcinoma, neuroblastoma, endometrial carcinoma, head and neck cancer or cervical cancer.

BRIEF DESCRIPTION OF GRAPHIC MATERIALS

The drawings form part of the description of the present invention and are included to further illustrate certain aspects of the embodiments described herein. Such embodiments may be better understood by reference to one or more of the following drawings, taken in conjunction with the detailed description.

Fig. 1 shows the in vitro inhibitory effect of an exemplary combination of the present invention, compound 24 with the PARP inhibitor olaparib, on MIA PaCa-2 cells.

Fig. 2 shows the in vitro inhibitory effect of an exemplary combination of the present invention, compound 24 with the PARP inhibitor BMN-673, on MDA-MB-231 cells.

Fig. 3 shows the in vitro inhibitory effect of an exemplary combination of the present invention, compound 24 with the EGFR inhibitor afatinib, on MIA PaCa-2 cells.

In Fig. 4the inhibitory effect was demonstratedin vitroillustrative combinations of the present invention, compound 24 with the SOS1 inhibitor BI-3406, against MIA PaCa-2 cells.

In Fig. 5the inhibitory effect was demonstratedin vitroillustrative combinations of the present invention, compound 24 with the EGFR inhibitor afatinib, in relation to MDA-MB-231 cells.

In Fig. 6the inhibitory effect was demonstratedin vitroillustrative combinations of the present invention, compound 24 with a CDK4/6 inhibitor, which is abemaciclib, in relation to MDA-MB-231 cells.

Fig. 7 is a line graph depicting the in vitro inhibitory effect of an exemplary combination of the present invention, compound 24, with the KRAS-G12C inhibitor AMG510 on MIA PaCa-2 cells.

Fig. 8 is a line graph depicting the in vivo combination efficacy of an exemplary combination of the present invention, Compound 24 with the EGFR inhibitor afatinib, in MIA PaCa-2 xenografts.

Fig. 9 is a line graph depicting the in vivo efficacy of an illustrative combination of the present invention, a combination of compound 24 with the SOS1 inhibitor BI-3406, against MIA PaCa-2 xenografts.

Fig. 10 is a line graph depicting the in vivo efficacy of an exemplary combination of the present invention, a combination of compound 24 with the ERK inhibitor ulixertinib (BVD-523), against MIA PaCa-2 xenografts.

DETAILED DESCRIPTION OF THE INVENTION

This description of illustrative embodiments is intended to be read in conjunction with the accompanying drawings/figures, which are to be considered as part of the entire written description. The drawings/figures are not necessarily drawn to scale, and certain features may be shown exaggerated in scale or somewhat schematically for purposes of clarity and emphasis.

Certain types of PRMT5 inhibitors, none of which are compounds of formula (I), can be combined with multiple modulators of cellular activity to treat cancer. Targeted therapeutics/modulators of cellular activity to treat cancer include PARP inhibitors (Cell Reports, 2018, 24, 2643–2657); EGFR inhibitors (Nat Cell Biol. 2011, Feb; 13(2): 174–181; Cancer Medicine, 2019, 8(5), 2414–2428); SOS1 inhibitors (Science, 2019, 363(6433), 1280–1281); RAF inhibitors (Sci Signal. 2011; 4(190): ra58); ERK inhibitors (Nat Cell Biol. 2011 Feb; 13(2):174–181); CDK4/6 inhibitors (Proc Natl Acad Sci USA. 2019; 116(36):17990–18000); MALT1 inhibitors (AACR Annual Meeting 2020; April 27–28, 2020 and June 22–24, 2020); BTK inhibitors (J Clin Oncol, 2020, 38(25): 2937); PI3K inhibitors (Leukemia. 2019, May 23. doi: 10.1038/s41375-019-0489-6); AKT inhibitors (J Cell Mol Med. 2019; 23(2):1333–1342); FGFR inhibitors (Cancer Res, 2015; 21(12); 2684–94); c-Met inhibitors (Sci Signal. 2011; 4(190): ra58; Cancer Medicine 2018; 7(3):869–882).

The present invention relates to a pharmaceutical combination comprising a combination of a PRMT5 inhibitor of formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist or an immune checkpoint inhibitor/modulator, wherein the PRMT5 inhibitor is a compound of formula (I),

its tautomeric form, its stereoisomer, its pharmaceutically acceptable salt, their polymorphic forms and their solvates,

Where

L ₁ is selected from -CR ^a R ^b -, -NR ^a -, S and O;

Z represents CH or N;

R ^a and R ^b are independently selected in each instance from hydrogen, substituted or unsubstituted alkyl, and substituted or unsubstituted cycloalkyl;

ring A is selected from

R ^c and R ^d are selected from substituted or unsubstituted alkyl or, together with the carbon atoms to which they are attached, form a C ₃ -C ₆ cycloalkyl ring;

R is selected from ^–NR4R5 , hydrogen, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted heteroaryl, and substituted or unsubstituted cycloalkyl ^;

R ¹ and R ² together with the carbon atoms to which they are attached form a bond to form –C=C- or R ¹ and R ² together with the carbon atoms to which they are attached form a cyclopropane ring;

R ^2' and R ^2a , which may be the same or different, are independently selected from hydrogen and substituted or unsubstituted alkyl;

R³independently selected at each occurrence from halogen, cyano, nitro, substituted or unsubstituted alkyl, -OR⁶, -NR⁷R⁸, substituted or unsubstituted cycloalkyl, -C(O)OH, -C(O)O-alkyl, -C(O)R⁹, -C(O)NR⁷R⁸, –NR⁷C(O)R⁹, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl and substituted or unsubstituted heterocyclyl;

R ⁴ and R ⁵ are independently selected from hydrogen, substituted or unsubstituted alkyl, and substituted or unsubstituted cycloalkyl;

R⁶ selected from hydrogen, substituted or unsubstituted alkyl, and substituted or unsubstituted cycloalkyl;

R ⁷ and R ⁸ are independently selected from hydrogen, substituted or unsubstituted alkyl, and substituted or unsubstituted cycloalkyl;

R ⁹ is selected from substituted or unsubstituted alkyl and substituted or unsubstituted cycloalkyl;

R¹⁰ selected from hydrogen, halogen and substituted or unsubstituted alkyl;

"n" is an integer between 0 and 4 inclusive;

if the alkyl or cycloalkyl group is substituted, it is substituted with 1-4 substituents independently selected from oxo (=O), halogen, cyano, cycloalkyl, aryl, heteroaryl, heterocyclyl, -OR ^7a , -C(=O)OH, -C(=O)O(alkyl), -NR ^8a R ^8b , -NR ^8a C(=O)R ^9a and –C(=O)NR ^8a R ^8b ;

if the aryl or heteroaryl group is substituted, it is substituted with 1-4 substituents independently selected from halogen, nitro, cyano, alkyl, haloalkyl, perhaloalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR^7a, -NR^8aR^8b, -NR^7aC(=O)R^9a, –C(=O)R^9a, –C(=O)NR^8aR^8b, -SO₂-alkyl, -C(=O)OH and -C(=O)O-alkyl;

if a heterocyclyl group is substituted, it is substituted at either a ring carbon atom or a ring heteroatom, and if substituted at a ring carbon atom, it is substituted by 1-4 substituents independently selected from oxo (=O), halogen, cyano, alkyl, cycloalkyl, perhaloalkyl, -OR^7a, –C(=O)NR^8aR^8b, -C(=O)OH, -C(=O)O-alkyl, -N(H)C(=O)(alkyl), -N(H)R^8aand -N(alkyl)₂; and if the heterocyclyl group is substituted at a nitrogen atom in the ring, then it is substituted with substituents independently selected from alkyl, cycloalkyl, aryl, heteroaryl, -SO₂(alkyl), –C(=O)R^9aAnd -C(=O)O(alkyl); if the heterocyclyl group is substituted at the sulfur atom in the ring, then it is substituted by 1 or 2 oxo groups (=O);

R ^7a is selected from hydrogen, alkyl, perhaloalkyl and cycloalkyl;

each of R ^8a and R ^8b is independently selected from hydrogen, alkyl and cycloalkyl; and

R ^9a is selected from alkyl and cycloalkyl.

In some embodiments, the present invention includes a pharmaceutical combination comprising a combination of a PRMT5 inhibitor of Formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator, wherein the PRMT5 inhibitor is:

(1S,2R,5R)-3-(2-(2-amino-3-bromoquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 1);

(1S,2R,5R)-3-(2-(2-amino-3-chloroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 2);

(1S,2R,5R)-3-(2-(2-amino-3-bromoquinolin-7-yl)ethyl)-5-(4-amino-5-fluoro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 3);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(((2-aminoquinolin-7-yl)thio)methyl)cyclopent-3-ene-1,2-diol (compound 4);

(1S,2R,5R)-3-(((2-amino-3-chloroquinolin-7-yl)thio)methyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 5);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(((2-aminoquinolin-7-yl)(methyl)amino)methyl)cyclopent-3-ene-1,2-diol (compound 6);

(1S,2R,5R)-3-(1-(2-amino-3-bromoquinolin-7-yl)propan-2-yl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compounds 7a and 7b);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(1-((2-(methylamino)quinolin-7-yl)oxy)ethyl)cyclopent-3-ene-1,2-diol (compounds 8a and 8b);

(1S,2R,5R)-3-(((2-amino-3-chloro-5-fluoroquinolin-7-yl)oxy)methyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylcyclopent-3-ene-1,2-diol (compound 9);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(2-(2-(methylamino)quinolin-7-yl)ethyl)cyclopent-3-ene-1,2-diol (compound 10);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(2-(3-methylimidazo[1,2-a]pyridin-7-yl)ethyl)cyclopent-3-ene-1,2-diol (compound 11);

(1S,2R,5R)-3-(((2-amino-3-chloro-5-fluoroquinolin-7-yl)oxy)methyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-methylcyclopent-3-ene-1,2-diol (compound 12);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 13);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-methyl-1H-pyrrolo[3,2-c]pyridin-1-yl)cyclopent-3-ene-1,2-diol hydrochloride (compound 14);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(1H-pyrrolo[3,2-c]pyridin-1-yl)cyclopent-3-ene-1,2-diol (compound 15);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 16);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(((2-aminoquinolin-7-yl)amino)methyl)cyclopent-3-ene-1,2-diol (compound 17);

(1S,2R,5R)-3-(((2-amino-3-chloro-5-fluoroquinolin-7-yl)oxy)methyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 18);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(((2-(methylamino)quinolin-7-yl)oxy)methyl)cyclopent-3-ene-1,2-diol (compound 19);

(1S,2R,5R)-3-(1-((2-amino-3-chloro-5-fluoroquinolin-7-yl)oxy)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 20a and 20b);

(1S,2R,5R)-3-(2-(2-amino-3-chloroquinolin-7-yl)ethyl)-5-(4-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 21);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(2-(2-(cyclobutylamino)quinolin-7-yl)ethyl)cyclopent-3-ene-1,2-diol (compound 22);

(1S,2R,5R)-3-(2-(2-amino-3-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 23);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 24);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-6-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 25);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-8-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 26);

(1S,2R,5R)-3-(2-(2-amino-3,3-dimethyl-3H-indol-6-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 27);

(1S,2R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(2-(2'-aminospiro[cyclobutan-1,3'-indol]-6'-yl)ethyl)cyclopent-3-ene-1,2-diol (compound 28);

(1S,2R,5R)-3-(2-(2-amino-3,5-dichloroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 29);

(1S,2R,5R)-3-(2-(2-amino-3-chloroquinolin-7-yl)ethyl)-5-(2-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 30);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 31);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 32);

(1S,2R,5R)-3-(1-(2-amino-3-chloro-5-fluoroquinolin-7-yl)propan-2-yl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compounds 33a and 33b);

(1S,2R,5R)-3-(1-(2-amino-3-chloroquinolin-7-yl)propan-2-yl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compounds 34a and 34b);

(1S,2R,5R)-3-(1-(2-amino-3-chloro-5-fluoroquinolin-7-yl)propan-2-yl)-5-(4-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compounds 35a and 35b);

(1S,2R,5R)-3-(1-(2-amino-3-chloro-5-fluoroquinolin-7-yl)propan-2-yl)-2-methyl-5-(4-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compounds 36a and 36b);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylcyclopent-3-ene-1,2-diol (compound 37);

(1S,2R,5R)-3-(1-(2-amino-3-chloro-5-fluoroquinolin-7-yl)propan-2-yl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-methylcyclopent-3-ene-1,2-diol (compound 38);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-2-methyl-5-(4-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 39);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-ethylcyclopent-3-ene-1,2-diol (compound 40);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-ethyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 41);

(1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 42);

(1S,2R,5R)-3-(2-(2-amino-3-bromo-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 43);

(1S,2R,5R)-3-(2-(2-amino-3-bromo-5-fluoroquinolin-7-yl)ethyl)-5-(4-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 44);

(1S,2R,5R)-3-(1-(2-amino-3-bromo-5-fluoroquinolin-7-yl)propan-2-yl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compounds 45a and 45b);

(1S,2R,5R)-3-(2-(2-amino-6-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 46);

(1S,2R,5R)-3-(2-(2-amino-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 47);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 48);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2-(methylamino)quinolin-7-yl)ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 49);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2-(isopropylamino)quinolin-7-yl)ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 50);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2-(cyclobutylamino)quinoline-7 -yl)ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 51);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2-((cyclopropylmethyl)amino)quinolin-7-yl)ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 52);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2-amino-8-fluoroquinoline- 7-yl)ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 53);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-methylquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 54);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-isopropylquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 55);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-(1,1-difluoroethyl)quinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 56);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-cyclopropylquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 57);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-methoxyquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 58);

2-amino-7-(2-((1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,3-dihydroxybicyclo[3.1.0]hexan-1-yl)ethyl)quinoline-3-carbonitrile (compound 59);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-fluoroquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d] pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 60);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-chloroquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 61);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-chloro-6-fluoroquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 62);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-chloro-8-fluoroquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 63);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-bromo-6-fluoroquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2, 3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 64);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(3-methylimidazo[1,2-a]pyridin-7-yl)ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 65);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3,3-dimethyl-3H-indol-6-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 66);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-4-(4-amino-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 67);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-chloro-6-fluoroquinolin-7-yl)ethyl)-4-(4-amino-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 68);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-4-(4-methyl-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 69);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-4-(7H-pyrrolo[2,3-d] pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 70);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2'-aminospiro[cyclobutan-1,3'-indol]-6'-yl)ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 71);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-bromo-5-fluoroquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2, 3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 72);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-bromoquinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d] pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 73);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2-aminoquinolin-7-yl)ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 74);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2-aminoquinazolin-7-yl)ethyl)bicyclo[3.1.0]hexane-2,3-diol (compound 75);

(1S,2R,3S,4R,5S)-1-((S)-1-(2-amino-3-bromoquinolin-7-yl)propan-2-yl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compounds 76a and 76b);

(1S,2R,3S,4R,5S)-1-((S)-2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)-1-cyclopropylethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compounds 77a and 77b);

(1S,2R,3S,4R,5S)-1-(1-(2-amino-3-chloro-5-fluoroquinolin-7-yl)propan-2-yl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compounds 78a and 78b);

(1R,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(2-(2-aminoquinolin-7-yl)propyl)bicyclo[3.1.0]hexane-2,3-diol (compounds 79a and 79b);

(1R,2R,3S,4R,5S)-1-(((2-amino-3-bromoquinolin-7-yl)oxy)methyl)-4-(4-amino-7H-pyrrolo[2,3-d ]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 80);

(1S,2R,3S,4R,5S)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(((2-aminoquinolin-7-yl)thio )methyl)bicyclo[3.1.0]hexane-2,3-diol (compound 81);

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-(4-fluorophenyl)quinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 82)

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-(pyridin-3-yl)quinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 83) or

(1R,2R,3S,4R,5S)-1-(2-(2-amino-3-(3-methylisoxazol-4-yl)quinolin-7-yl)ethyl)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)bicyclo[3.1.0]hexane-2,3-diol (compound 84) and

their pharmaceutically acceptable salts.

In some embodiments, the pharmaceutical combination comprises a compound that is (1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (Compound 24) and at least one targeted agent/modulator of cellular activity that is an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, FGFR, PRMT type I inhibitor, STING agonist or immune checkpoint inhibitor/modulator, for use in the treatment and/or prevention of cancer.

In some embodiments, the pharmaceutical combination comprises at least one PRMT5 inhibitor of Formula 1 and an inhibitor of EGFR or oncogenic variants thereof. In some embodiments, the EGFR inhibitor is afatinib, (( E ) -N- [4-(3-chloro-4-fluoroanilino)-7-[( 3S )-oxolan-3-yl]oxyquinazolin-6-yl]-4-(dimethylamino)but-2-enamide), osimertinib, ( N- [2-[2-(dimethylamino)ethyl-methylamino]-4-methoxy-5-[[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino]phenyl]prop-2-enamide), erlotinib, ( N- (3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine), and gefitinib, ( N -(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholin-4-ylpropoxy)quinazolin-4-amine):

(afatinib),

(osimertinib),

(erlotinib)

or

(gefitinib).

In some embodiments, the pharmaceutical combination comprises at least one PRMT5 inhibitor of formula 1 and an SOS1 inhibitor. In some embodiments, the SOS1 inhibitor is BI-3406 (N-((R)-1-(3-amino-5-(trifluoromethyl)phenyl)ethyl)-7-methoxy-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-amine):

In some embodiments, the pharmaceutical combination comprises at least one PRMT5 inhibitor of formula 1 and an ERK inhibitor. In some embodiments, the ERK inhibitor is ulixertinib, ( N -[( 1S )-1-(3-chlorophenyl)-2-hydroxyethyl]-4-[5-chloro-2-(propan-2-ylamino)pyridin-4-yl]-1H - pyrrole-2-carboxamide), MK-8353, ((S)-N-(3-(6-isopropoxypyridin-3-yl)-1H-indazol-5-yl)-1-(2-(4-(4-(1-methyl-1H-1,2,4-triazol-3-yl)phenyl)-3,6-dihydropyridin-1(2H)-yl)-2-oxoethyl)-3-(methylthio)pyrrolidine-3-carboxamide), and ravoxertinib, (1-[(1S)-1-(4-chloro-3-fluorophenyl)-2-hydroxyethyl]-4-[2-[(2-methylpyrazol-3-yl)amino]pyrimidin-4-yl]pyridin-2-one):

(ulixertinib),

(MK-8353)

or

(ravoxertinib).

In some embodiments, the pharmaceutical combination comprises at least one PRMT5 inhibitor of Formula 1 and a PARP inhibitor. In some embodiments, the PARP inhibitor is olaparib, (4-[[3-[4-(cyclopropanecarbonyl)piperazine-1-carbonyl]-4-fluorophenyl]methyl]-2H-phthalazin-1-one), niraparib, ((S)-2-(4-(piperidin-3-yl)phenyl)-2H-indazole-7-carboxamide), rucaparib, (6-fluoro-2-[4-(methylaminomethyl)phenyl]-3,10-diazatricyclo[6.4.1.04,13]trideca-1,4,6,8(13)-tetraen-9-one), and talazoparib (BMN-673), ((11S,12R)-7-fluoro-11-(4-fluorophenyl)-12-(2-methyl-1,2,4-triazol-3-yl)-2,3,10-triazatricyclo[7.3.1.05,13]trideca-1,5(13),6,8-tetraen-4-one):

(olaparib),

(niraparib),

(rukaparib)

or

(talazoparib).

In some embodiments, the pharmaceutical combination comprises at least one PRMT5 inhibitor of formula 1 and a CDK4/6 inhibitor. In some embodiments, the CDK4/6 inhibitor is palbociclib, (6-acetyl-8-cyclopentyl-5-methyl-2-[(5-piperazin-1-ylpyridin-2-yl)amino]pyrido[2,3-d]pyrimidin-7-one), ribociclib, (7-cyclopentyl-N,N-dimethyl-2-[(5-piperazin-1-ylpyridin-2-yl)amino]pyrrolo[2,3-d]pyrimidine-6-carboxamide), and abemaciclib, (N-[5-[(4-ethylpiperazin-1-yl)methyl]pyridin-2-yl]-5-fluoro-4-(7-fluoro-2-methyl-3-propan-2-ylbenzimidazol-5-yl)pyrimidin-2-amine):

(palbociclib),

(ribociclib)

or

(abemaciclib).

In some embodiments, the pharmaceutical combination comprises at least one PRMT5 inhibitor of formula 1 and a BTK inhibitor. In some embodiments, the BTK inhibitor is ibrutinib, (1-[(3R)-3-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]piperidin-1-yl]prop-2-en-1-one), zanubrutinib, (7S)-2-(4-phenoxyphenyl)-7-(1-prop-2-enoylpiperidin-4-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide), or acalabrutinib, (4-[8-amino-3-[(2S)-1-but-2-inoylpyrrolidin-2-yl]imidazo[1,5-a]pyrazin-1-yl]-N-pyridin-2-ylbenzamide):

(ibrutinib),

(zanubrutinib)

or

(acalabrutinib).

In some embodiments, the present invention includes a pharmaceutical combination that comprises (a) a compound that is (1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol, or a pharmaceutically acceptable salt thereof, and (b) afatinib, for use in the treatment and/or prevention of cancer in a subject in need thereof.

In some embodiments, the present invention includes a pharmaceutical combination comprising (a) a compound that is (1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol, or a pharmaceutically acceptable salt thereof, and (b) BI-3406, for use in the treatment and/or prevention of cancer in a subject in need thereof.

In some embodiments, the present invention includes a pharmaceutical combination comprising (a) a compound that is (1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol, or a pharmaceutically acceptable salt thereof, and (b) ulixertinib, for use in the treatment and/or prevention of cancer in a subject in need thereof.

In some embodiments, the present invention includes a pharmaceutical combination comprising a PRMT5 inhibitor of formula (I) with an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator. The PRMT5 inhibitor of formula (I) can be administered simultaneously, in parallel, sequentially, one after the other, alternately or separately with at least one targeted agent/modulator of cellular activity (for example, the administration of the PRMT5 inhibitor of formula (I) and at least one targeted agent/modulator of cellular activity can be performed in any sequence).

In some embodiments, the present invention includes a method of treating and/or preventing cancer in a human subject in need thereof, the method comprising administering to the human subject a therapeutically effective amount of a pharmaceutical combination comprising a PRMT5 inhibitor of Formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator, defined herein, thereby treating and/or preventing cancer in a human subject.

In some embodiments, the present invention includes a method of treating and/or preventing the development of cancer in a human subject, the method comprising administering to the human subject a therapeutically effective amount of a pharmaceutical combination comprising a PRMT5 inhibitor of Formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator as defined in in this document, together with at least one of a pharmaceutically acceptable carrier and a pharmaceutically acceptable diluent, thereby treating and/or preventing cancer in a human subject.

In some embodiments, the present invention includes a pharmaceutical combination comprising a PRMT5 inhibitor of Formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator as defined herein, for use in the treatment and/or prevention of cancer in a subject in need thereof.

In some embodiments, the present invention includes a pharmaceutical combination comprising a PRMT5 inhibitor of Formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator as defined herein, for use in the treatment of cancer in a subject in need thereof.

In some embodiments, the present invention includes a pharmaceutical combination comprising a PRMT5 inhibitor of Formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator as defined herein, for use in reducing the rate of tumor growth, reducing the size of a tumor, or reducing one or more symptoms associated with the presence of a tumor in the subject requiring it.

In some embodiments, the present invention includes a pharmaceutical combination comprising a PRMT5 inhibitor of Formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator, which can be used to treat and/or prevent various cancers, which may include or exclude: multiforme Glioblastoma, prostate cancer, pancreatic cancer, mantle cell lymphoma, non-Hodgkin's lymphomas and diffuse large B-cell lymphoma, acute myeloid leukemia, acute lymphoblastic leukemia, multiple myeloma, non-small cell lung cancer, small cell lung cancer, breast cancer, triple-negative breast cancer, gastric cancer, colorectal cancer, ovarian cancer, bladder cancer, hepatocellular carcinoma, melanoma, sarcoma, oropharyngeal squamous cell carcinoma, chronic myelogenous leukemia, epidermal squamous cell carcinoma, nasopharyngeal carcinoma, neuroblastoma, endometrial carcinoma, head and neck cancer and cervical cancer.

Definitions

General terms used in the formula may be defined as set forth below; however, the meaning set forth should not be interpreted as limiting the scope of the term per se .

As used herein, the indication of a numerical range for a variable is intended to express that the present invention can be practiced with the variable equal to any value within the range. Therefore, with a variable that is inherently discrete, the variable may be equal to any integer value belonging to the numerical range, including the endpoints of the range. Similarly, with a variable that is inherently continuous, the variable may be equal to any real value in the numerical range, including the endpoints of the range. As an example, a variable that is described as taking values from 0 to 2 may represent 0, 1, or 2 in the case of variables that are inherently discrete, and may represent 0.0, 0.1, 0.01, 0.001, or any other real value in the case of variables that are inherently continuous.

As used in this document, the term "approximately" is intended to quantify the numerical values it modifies, designating such values as variables that are within an acceptable margin of error. Unless a specific margin of error, such as a standard deviation from a mean, is specified, the term "approximately" means plus or minus 10% of the numerical value of the number to which it applies. Therefore, for example, approximately 50% means in the range of 45% to 55%.

As used in this document, the terms "halogen" or "halo" mean fluorine, chlorine, bromine or iodine.

As used herein, the term "alkyl" means a hydrocarbon radical derived from an alkane that contains exclusively carbon and hydrogen atoms in the main chain, does not include any unsaturation, contains from one to six carbon atoms and is attached to the rest of the molecule by a single bond, such as (C₁-C₆)alkyl or (C₁-C₄)alkyl, illustrative groups include, for example, methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, etc. Unless otherwise stated or listed, all alkyl groups described or claimed herein may be straight chain or branched chain.

As used herein, the term "alkenyl" means a hydrocarbon radical containing 2-10 carbon atoms and having at least one carbon-carbon double bond. Non-limiting examples of alkenyl groups include, for example, (C₂-C₆)alkenyl, (C₂-C₄)alkenyl, ethenyl, 1-propenyl, 2-propenyl (allyl), isopropenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, etc. Unless otherwise stated or listed, all alkenyl groups described or claimed herein may be straight chain or branched chain.

As used herein, the term "alkynyl" means a hydrocarbon radical containing 2-10 carbon atoms and providing at least one carbon-carbon triple bond. Non-limiting examples of alkynyl groups include, for example, ( _C2 - _C6 )alkynyl, ( _C2 - _C4 )alkynyl, ethynyl, propynyl, butynyl, and the like. Unless otherwise noted or listed, all alkynyl groups described or claimed herein may be straight chain or branched chain.

As used herein, the term "haloalkyl" means an alkyl group as defined above that is substituted with one or more halogen atoms as defined above. For example, means (C ₁ -C ₆ )haloalkyl or (C ₁ -C ₄ )haloalkyl. Accordingly, haloalkyl can be monohaloalkyl, dihaloalkyl or polyhaloalkyl, including perhaloalkyl. Monohaloalkyl can contain one iodine, bromine, chlorine or fluorine atom. Dihaloalkyl and polyhaloalkyl groups can be substituted with two or more atoms of a single halogen or a combination of atoms of different halogens. Accordingly, polyhaloalkyl is substituted with no more than 12 halogen atoms. Non-limiting examples of haloalkyl include fluoromethyl, difluoromethyl, trifluoromethyl, chloromethyl, dichloromethyl, trichloromethyl, pentafluoroethyl, heptafluoropropyl, difluorochloromethyl, dichlorofluoromethyl, difluoroethyl, difluoropropyl, dichloroethyl, dichloropropyl, and the like. Perhaloalkyl means an alkyl in which all hydrogen atoms are replaced by halogen atoms. Unless otherwise noted or listed, all haloalkyl groups described or claimed herein may be straight chain or branched chain.

As used herein, the term "alkoxy" means an alkyl group attached via _an oxygen bond to the rest of the molecule. Typical examples of such groups are _-OCH3 and _-OC2H5 . Unless otherwise noted or listed, all alkoxy groups described or claimed herein may be straight chain or branched chain.

As used herein, the term "alkoxyalkyl" means an alkoxy group, as defined above, directly linked to an alkyl group, as defined above, for example, -CH₂-O-CH₃, -CH₂-O-CH₂CH₃, -CH₂CH₂-O-CH₃etc.

As used herein, the term "cycloalkyl" means a non-aromatic mono- or polycyclic ring system containing 3-12 carbon atoms, such as (C₃-C₁₀)cycloalkyl, (C₃-C₆)cycloalkyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc. Examples of polycyclic cycloalkyl groups include, but are not limited to, perhydronaphthyl, adamantyl and norbornyl groups, bridged cyclic groups or spirobicyclic groups, for example, spiro(4,4)non-2-yl, etc.

The term "aryl" as used herein means an aromatic radical containing 6-14 carbon atoms, including monocyclic, bicyclic and tricyclic aromatic systems such as phenyl, naphthyl, tetrahydronaphthyl, indanyl and biphenyl, etc.

As used herein, the terms "heterocyclic ring" or "heterocyclyl ring" or "heterocyclyl" unless otherwise specified, mean a substituted or unsubstituted non-aromatic 3-15 membered ring that contains carbon atoms and one or more heteroatoms independently selected from N, O or S. The heterocyclic ring may be a mono-, bi- or tricyclic ring system, which may include fused, bridged or spiro ring systems, and the nitrogen, carbon, oxygen or sulfur atoms in the heterocyclic ring may optionally be oxidized to various oxidation states. In addition, the nitrogen atom may be optionally quaternized, the heterocyclic ring or heterocyclyl may optionally contain one or more olefinic bonds, and one or two carbon atoms in the heterocyclic ring or heterocyclyl may be separated by _-CF2- , -C(O)-, -S(O)-, S(O) ₂ , etc. In addition, the heterocyclic ring may also be condensed with an aromatic ring. Non-limiting examples of heterocyclic rings include azetidinyl, benzopyranyl, chromanyl, decahydroisoquinolyl, indolinyl, isoindolinyl, isochromanyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, oxazolinyl, oxazolidinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, octahydroindolyl, octahydroisoindolyl, perhydroazepinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, piperidinyl, phenothiazinyl, phenoxazinyl, quinuclidinyl, tetrahydroisoquinolyl, tetrahydrofuryl, tetrahydropyranyl, thiazolinyl, thiazolidinyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfonindoline, benzodioxole, tetrahydroquinoline, tetrahydrobenzopyran, etc. The heterocyclic ring can be attached to any atom of the heterocyclic ring, resulting in the formation of a stable structure.

As used herein, unless otherwise specified, the term "heteroaryl" means a substituted or unsubstituted 5- to 14-membered aromatic heterocyclic ring with one or more heteroatoms, independently selected from N, O or S. Heteroaryl can be a mono-, bi- or tricyclic ring system. The heteroaryl ring can be attached at any atom of the heteroaryl ring, resulting in a stable structure. Non-limiting examples of a heteroaryl ring include oxazolyl, isoxazolyl, imidazolyl, furyl, indolyl, isoindolyl, pyrrolyl, triazolyl, triazinyl, tetrazolyl, thienyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzofuranyl, benzothiazolyl, benzoxazolyl, benzimidazolyl, benzothienyl, carbazolyl, quinolinyl, isoquinolinyl, quinazolinyl, cinnolinyl, naphthyridinyl, pteridinyl, purinyl, quinoxalinyl, quinolyl, isoquinolyl, thiadiazolyl, indolizinyl, acridinyl, phenazinyl, phthalazinyl, and the like.

The compounds of the present invention may have one or more chiral centers. The absolute stereochemistry at each chiral center may be "R" or "S". The compounds of the present invention include all diastereomers and enantiomers and mixtures thereof. Unless otherwise specifically indicated, reference to one stereoisomer is used to refer to any of the possible stereoisomers. Whenever the stereoisomeric composition is not specified, it is understood that all possible stereoisomers are included.

As used herein, the term "stereoisomer" means a compound composed of the same atoms linked by the same bonds but having different three-dimensional structures that are not interchangeable. The three-dimensional structures are called configurations. As used herein, the term "enantiomer" means two stereoisomers whose molecules are non-superimposable mirror images of each other. The term "chiral center" means a carbon atom to which four different groups are attached. As used herein, the term "diastereomers" means stereoisomers that are not enantiomers. The terms "racemate" or "racemic mixture" mean a mixture of equal parts of enantiomers.

As used herein, the term "tautomer" means a compound that undergoes rapid proton transfers from one atom of the compound to another atom of the compound. Some compounds described herein may exist as tautomers with different hydrogen attachment points. Individual tautomers as well as mixtures thereof are encompassed by the compounds of formula (I).

As used herein, the expressions "treating" or "treating" a condition, disorder, or pathological condition include: (a) preventing or slowing the onset of clinical symptoms of the condition, disorder, or pathological condition developing in a subject who may have or be predisposed to the condition, disorder, or pathological condition but does not yet experience or exhibit clinical or subclinical symptoms of the condition, disorder, or pathological condition; (b) suppressing the condition, disorder, or pathological condition, i.e., arresting or slowing the progression of the disease or at least one of its clinical or subclinical symptoms; (c) alleviating the severity of the disease, disorder, or pathological condition or at least one of its clinical or subclinical symptoms; or (d) alleviating the disease, i.e., causing regression or reduction in the intensity of the condition, disorder, or pathological condition or at least one of its clinical or subclinical symptoms.

As used herein, the terms "treat," "ameliorate," and "suppress," and related words used herein, do not necessarily mean 100% or complete cure, amelioration, or suppression. Rather, there are varying degrees of treatment, amelioration, and suppression that are known to one of ordinary skill in the art to have potential benefit or therapeutic effect. In this regard, the disclosed methods can provide any degree of any level of treatment, amelioration, or suppression of a disorder in a mammal. For example, a disorder, including its symptoms or pathological conditions, can be reduced by, for example, 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, or 10%. Furthermore, the treatment, amelioration, or suppression provided by the method of the present invention can include treatment, amelioration, or suppression of one or more conditions or symptoms of the disorder, such as cancer. Also, for the purposes of this document, "treatment," "alleviation," or "suppression" may include delaying the onset of a disorder or a symptom or condition thereof.

As used herein, the term "inhibitor" means a molecule that binds to an enzyme to inhibit the activity of said enzyme either partially or completely.

As used herein, the terms "patient" and "subject" are used interchangeably and may be used to refer to any living organism that can be treated with the pharmaceutical combination disclosed herein. The terms "patient" and "subject" include mammals (such as a primate or a human) and other animals, such as domestic animals (e.g., pets, including cats and dogs) and non-domesticated animals (such as wild animals).

As used herein, the term "therapeutically effective amount" means an amount of a compound that, when administered to a subject for the treatment of a disease, disorder, or condition, is sufficient to provide the effect in the subject that is the purpose of the administration. "Therapeutically effective amount" will vary depending on the compound, the route of administration, the disease and its severity, and the age, weight, physical condition, and responsiveness of the subject being treated.

As used herein, the term "pharmaceutically acceptable carrier" refers to an ingredient in a pharmaceutical formulation other than the active ingredient that can be safely administered to a subject. A pharmaceutically acceptable carrier includes, but is not limited to, a buffer, an excipient, a stabilizer, or a preservative. Suitable carriers and diluents include buffered, aqueous solutions, saline, dextrose, glycerol, isotonic saline solutions such as phosphate buffered saline, isotonic water, and the like, and combinations thereof. In some embodiments, carriers may include propylene glycol, dimethyl isosorbide, and water, and even more particularly phosphate buffered saline, isotonic water, deionized water, monofunctional alcohols, and symmetrical alcohols.

As used herein, the term "tumor" refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all precancerous and cancerous cells and tissues. The terms "cancer," "cancerous," "cell proliferative disorder," "proliferative disorder," and "tumor" are not mutually exclusive as defined herein.

As used herein, the phrase "pharmaceutically acceptable salt" refers to pharmaceutically acceptable organic or inorganic salts of a compound of the present invention. Illustrative salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate "mesylate", ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e. 1,1′-methylene-bis(2-hydroxy-3-naphthoate) salts. A pharmaceutically acceptable salt may include another molecule, such as an acetate ion, a succinate ion, or another counterion. In some embodiments, the counterion is any organic or inorganic moiety that stabilizes the charge on the parent compound. In addition, a pharmaceutically acceptable salt may contain more than one charged atom in its structure. In cases where multiple charged atoms are part of a pharmaceutically acceptable salt, there may be multiple counterions for it. Thus, a pharmaceutically acceptable salt may contain one or more charged atoms and/or one or more counterions.

In some embodiments, if the compound of the present invention is a base, the desired pharmaceutically acceptable salt is prepared by any suitable method available in the art, for example, treating the free base with an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, methanesulfonic acid, phosphoric acid, and the like, or an organic acid such as acetic acid, trifluoroacetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a pyranosidyl acid such as glucuronic acid or galacturonic acid, an alpha hydroxy acid such as citric acid or tartaric acid, an amino acid such as aspartic acid or glutamic acid, an aromatic acid such as benzoic acid or cinnamic acid, a sulfonic acid an acid such as p-toluenesulfonic acid or ethanesulfonic acid or the like.

In some embodiments, if the compound of the present invention is an acid, the desired pharmaceutically acceptable salt is prepared by any suitable method, such as treating the free acid with an inorganic or organic base such as an amine (primary, secondary or tertiary), an alkali metal hydroxide or an alkaline earth metal hydroxide, or the like. Illustrative examples of suitable salts include, but are not limited to, organic salts derived from amino acids such as glycine and arginine, ammonia, primary, secondary and tertiary amines and cyclic amines such as piperidine, morpholine and piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium.

As used herein, the term "solvate" refers to an association or complex consisting of one or more solvent molecules and a compound of the present invention. Examples of solvents with which solvates are formed include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine.

Compositions

In some embodiments, EGFR inhibitors that may be used in combination with the PRMT5 inhibitors of Formula (I) may include or exclude afatinib, osimertinib, erlotinib, or gefitinib, or any other agent that inhibits the activity of EGFR enzymes or oncogenic variants thereof.

In some embodiments, CDK4/6 inhibitors that may be used in combination with PRMT5 inhibitors of Formula (I) may include or exclude palbociclib, ribociclib, or abemaciclib, or any other agent that inhibits the activity of CDK4 and CDK6 enzymes.

In some embodiments, RAF inhibitors that may be used in combination with the PRMT5 inhibitors of Formula (I) may include or exclude dabrafenib, regorafenib, encorafenib, or inhibitors of all forms of RAF, such as RAF265, or any other agent that inhibits the activity of RAF isoforms (ARAF, BRAF, and CRAF).

In some embodiments, ERK inhibitors that may be used in combination with the PRMT5 inhibitors of Formula (I) may include or exclude ulixertinib, MK-8353, or ravoxertinib, or any other agent that inhibits the activity of various ERK1/2 kinases.

In some embodiments, SOS1 inhibitors that may be used in combination with PRMT5 inhibitors of Formula (I) may include or exclude BI-3406.

In some embodiments, MALT1 inhibitors that may be used in combination with PRMT5 inhibitors of Formula (I) may include or exclude MI-2 or JNJ-67856633.

In some embodiments, BTK inhibitors that may be used in combination with PRMT5 inhibitors of Formula (I) may include or exclude ibrutinib, zanubrutinib, or acalabrutinib.

In some embodiments, PI3K inhibitors that may be used in combination with the PRMT5 inhibitors of Formula (I) may include or exclude alpelisib, copanlisib, duvelisib, BEZ-235, gedatolisib, buparlisib, or agents that inhibit the activity of one or more PI3K isoforms (α, β, δ, and γ), or dual PI3K-mTOR inhibitors.

In some embodiments, AKT inhibitors that may be used in combination with PRMT5 inhibitors of Formula (I) may include or exclude GSK690693, AZD5363, or ipatasertib.

In some embodiments, FGFR inhibitors that may be used in combination with PRMT5 inhibitors of Formula (I) may include or exclude dovitinib, AZD4547, BGJ398, or JNJ 42756493.

In some embodiments, c-Met inhibitors that may be used in combination with the PRMT5 inhibitors of Formula (I) may include or exclude tivantinib, cabozantinib, crizotinib, or capmatinib.

In some embodiments, KRAS inhibitors and KRAS-G12C inhibitors that may be used in combination with PRMT5 inhibitors of Formula (I) may include or exclude sotorasib (AMG510), JNJ74699157, or GDC-6036.

In some embodiments, MEK inhibitors that may be used in combination with PRMT5 inhibitors of Formula (I) may include or exclude cobimetinib, trametinib, SF-2626.

In some embodiments, Bcl-2 inhibitors that may be used in combination with PRMT5 inhibitors of Formula (I) may include or exclude navitoclax, venetoclax, AZD-4320, alvocidib hydrochloride, TP-1287, APG-2575.

In some embodiments, PRMT type I inhibitors that may be used in combination with PRMT5 inhibitors of Formula (I) may include or exclude GSK-3368715.

In some embodiments, MAT2A inhibitors that may be used in combination with PRMT5 inhibitors of Formula (I) may include or exclude AG24512 or AG25696.

In some embodiments, STING agonists that may be used in combination with the PRMT5 inhibitors of Formula (I) may include or exclude MK-1454, GSK-532, SYNB-1891, E-7766, or BMS-986301.

In some embodiments, immune checkpoint inhibitors/modulators (immuno-oncology agents) that can be used in combination with the PRMT5 inhibitors of Formula (I) can include or exclude nivolumab, ipilimumab, pembrolizumab, omburtamab, tremelimumab, pidilizumab, varlilumab, epacadostat, CT-011, AMP-224, and MDX-1106. Checkpoint inhibitors are agents that target checkpoint proteins or derivatives thereof and can be referred to as "checkpoint inhibitors." Checkpoint inhibitors can include or exclude proteins, polypeptides, amino acid residues, and monoclonal or polyclonal antibodies. PRMT5 inhibitor compounds can be administered in combination with one or more checkpoint inhibitors. Checkpoint inhibitors can bind, for example, to ligands or proteins that are found in any of the CD28/CTLA-4-like family of T cell regulators. Targets for checkpoint inhibitors include, but are not limited to, receptors or co-receptors (e.g., CTLA-4; CD8) expressed on effector or regulatory cells of the immune system (e.g., T cells); proteins expressed on the surface of antigen-presenting cells (e.g., those expressed on the surface of activated T cells, including PD-1, PD-2, PD-L1 PD-L2, 4-1BB, and OX40); metabolic enzymes or metabolic enzymes expressed by both tumors and tumor-infiltrating immune cells (e.g., indoleamine (IDO), including isoforms such as IDO1 and IDO2); proteins that belong to the immunoglobulin superfamily (e.g., lymphocyte activation gene 3, also known as LAG3); proteins that belong to the B7 superfamily (e.g., B7-H3 or its homologues). B7 proteins can be found on both activated antigen-presenting cells and T cells.

Methods

In some embodiments, the present invention provides the use of a pharmaceutical combination comprising a PRMT5 inhibitor of Formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for use in the treatment of a disease or pathological condition in which Inhibition of PRMT5 protein has a beneficial effect.

In some embodiments, the present invention provides a method of treating and/or preventing cancer in a human subject in need thereof, the method comprising administering to the human subject a pharmaceutical combination comprising a PRMT5 inhibitor composition of Formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator. answer. The disease that is cancer may be glioblastoma multiforme, prostate cancer, pancreatic cancer, mantle cell lymphoma, non-Hodgkin lymphomas and diffuse large B-cell lymphoma, acute myeloid leukemia, acute lymphoblastic leukemia, multiple myeloma, non-small cell lung cancer, small cell lung cancer, breast cancer, triple-negative breast cancer, gastric cancer, colorectal cancer, ovarian cancer, bladder cancer, hepatocellular carcinoma, melanoma, sarcoma, oropharyngeal squamous cell carcinoma, chronic myelogenous leukemia, epidermal squamous cell carcinoma, nasopharyngeal carcinoma, neuroblastoma, or endometrial carcinoma, or cervical cancer.

In some embodiments, the present invention provides a method for treating and/or preventing cancer in a subject in need thereof. In some embodiments, the present invention provides a method for slowing the rate of tumor growth, reducing the size of a tumor, reducing one or more symptoms associated with the presence of a tumor in a subject in need thereof, or eliminating a tumor. Cancer may also include or exclude solid tumors, various types of leukemia or lymphoma.

Introduction

In some embodiments, the pharmaceutical combination provided herein may comprise a combination of a PRMT5 inhibitor of formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator, and administered by any route appropriate to the pathological condition to be treated. Suitable routes may include or exclude oral, parenteral (including subcutaneous, intramuscular, intravenous, intraarterial, intradermal, intrathecal and epidural), intraperitoneal (IP), transdermal, rectal, nasal, topical (including buccal and sublingual), vaginal, intrapulmonary and intranasal. In some embodiments, when performing topical treatment, the compounds are administered by intratumoral administration, including perfusion, or by otherwise contacting the tumor with the inhibitor. It is understood that the preferred route may vary, for example, depending on the condition of the recipient/subject. In some embodiments, where the compound is administered orally, it is formulated as a pill, capsule, tablet, etc., with a pharmaceutically acceptable carrier and/or excipient. In some embodiments where the compound is administered parenterally, it is formulated with a pharmaceutically acceptable carrier for parenteral administration and as a unitary injection dosage form as described herein.

Suitable doses and dosage regimens can be determined using therapeutic dose range determination techniques that depend on the subject's body weight, body volume, body surface area, body sites for administration, routes of administration, and prognosis for the stage of cancer in the subject. Typically, treatment is initiated with lower dosages that are less than the optimal dose of the composition of the present invention. The dosage is then increased in small increments until the optimal effect under the circumstances is achieved. The method of the present invention may involve the administration of about 0.1 μg to about 50 mg of at least one compound of the present invention per kg of body weight of the subject. For example, for a patient weighing 70 kg, dosages of about 10 μg to about 200 mg of the compound may be used, depending on the physiological response of the patient.

In some embodiments, the dose of the pharmaceutically active agents described herein for the methods of treating and/or preventing cancer described herein can be from about 0.001 to about 1 mg/kg of subject body weight per day, such as about 0.001 mg, 0.002 mg, 0.005 mg, 0.010 mg, 0.015 mg, 0.020 mg, 0.025 mg, 0.050 mg, 0.075 mg, 0.1 mg, 0.15 mg, 0.2 mg, 0.25 mg, 0.5 mg, 0.75 mg, or 1 mg/kg of body weight per day. The dosage of the pharmaceutically active agent(s) described herein for the disclosed methods may be from about 1 to about 1000 mg/kg body weight of the subject to be treated per day, such as about 1 mg, 2 mg, 5 mg, 10 mg, 15 mg, 0.020 mg, 25 mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 500 mg, 750 mg, or 1000 mg/kg body weight per day.

In some embodiments, a dose of a PRMT5 inhibitor of Formula (I) in a pharmaceutical combination for the methods of treating and/or preventing cancer described herein can be administered at doses of 1 mg/kg per day, about 2 mg/kg per day, about 5 mg/kg per day, about 10 mg/kg per day, about 15 mg/kg per day, about 20 mg/kg per day, about 25 mg/kg per day, about 30 mg/kg per day, about 35 mg/kg per day, about 40 mg/kg per day, about 45 mg/kg per day, about 50 mg/kg per day, about 60 mg/kg per day, about 70 mg/kg per day, about 80 mg/kg per day, about 90 mg/kg per day, about 100 mg/kg per day, about 125 mg/kg per day, about 150 mg/kg per day, about 175 mg/kg per day, approximately 200 mg/kg per day, approximately 250 mg/kg per day, or approximately 300 mg/kg per day, in combination with an EGFR inhibitor that may be administered at doses of approximately 1 mg/kg per day, approximately 2 mg/kg per day, approximately 5 mg/kg per day, approximately 10 mg/kg per day, approximately 15 mg/kg per day, approximately 20 mg/kg per day, approximately 25 mg/kg per day, approximately 30 mg/kg per day, approximately 35 mg/kg per day, approximately 40 mg/kg per day, approximately 45 mg/kg per day, approximately 50 mg/kg per day, approximately 60 mg/kg per day, approximately 70 mg/kg per day, approximately 80 mg/kg per day, approximately 90 mg/kg per day, approximately 100 mg/kg per day, approximately 125 mg/kg per day, approximately 150 mg/kg per day, approximately 175 mg/kg per day, approximately 200 mg/kg per day, approximately 250 mg/kg per day, or approximately 300 mg/kg per day.

In some embodiments, a dose of a PRMT5 inhibitor of Formula (I) in a pharmaceutical combination for the methods of treating and/or preventing cancer described herein can be administered at doses of 1 mg/kg per day, about 2 mg/kg per day, about 5 mg/kg per day, about 10 mg/kg per day, about 15 mg/kg per day, about 20 mg/kg per day, about 25 mg/kg per day, about 30 mg/kg per day, about 35 mg/kg per day, about 40 mg/kg per day, about 45 mg/kg per day, about 50 mg/kg per day, about 60 mg/kg per day, about 70 mg/kg per day, about 80 mg/kg per day, about 90 mg/kg per day, about 100 mg/kg per day, about 125 mg/kg per day, about 150 mg/kg per day, about 175 mg/kg per day, approximately 200 mg/kg per day, approximately 250 mg/kg per day, or approximately 300 mg/kg per day, in combination with an SOS1 inhibitor that may be administered at doses of approximately 1 mg/kg per day, approximately 2 mg/kg per day, approximately 5 mg/kg per day, approximately 10 mg/kg per day, approximately 15 mg/kg per day, approximately 20 mg/kg per day, approximately 25 mg/kg per day, approximately 30 mg/kg per day, approximately 35 mg/kg per day, approximately 40 mg/kg per day, approximately 45 mg/kg per day, approximately 50 mg/kg per day, approximately 60 mg/kg per day, approximately 70 mg/kg per day, approximately 80 mg/kg per day, approximately 90 mg/kg per day, approximately 100 mg/kg per day, approximately 125 mg/kg per day, approximately 150 mg/kg per day, approximately 175 mg/kg per day, approximately 200 mg/kg per day, approximately 250 mg/kg per day, or approximately 300 mg/kg per day.

In some embodiments, a dose of a PRMT5 inhibitor of Formula (I) in a pharmaceutical combination for methods of treating and/or preventing cancer as described herein for methods of treating and/or preventing a disease or condition as described above with respect to PRMT5 can be administered at doses of 1 mg/kg per day, about 2 mg/kg per day, about 5 mg/kg per day, about 10 mg/kg per day, about 15 mg/kg per day, about 20 mg/kg per day, about 25 mg/kg per day, about 30 mg/kg per day, about 35 mg/kg per day, about 40 mg/kg per day, about 45 mg/kg per day, about 50 mg/kg per day, about 60 mg/kg per day, about 70 mg/kg per day, about 80 mg/kg per day, about 90 mg/kg per day, about 100 mg/kg per day, about 125 mg/kg per day, or about 150 mg/kg per day. day, approximately 150 mg/kg per day, approximately 175 mg/kg per day, approximately 200 mg/kg per day, approximately 250 mg/kg per day, or approximately 300 mg/kg per day, in combination with an ERK inhibitor that may be administered at doses of approximately 1 mg/kg per day, approximately 2 mg/kg per day, approximately 5 mg/kg per day, approximately 10 mg/kg per day, approximately 15 mg/kg per day, approximately 20 mg/kg per day, approximately 25 mg/kg per day, approximately 30 mg/kg per day, approximately 35 mg/kg per day, approximately 40 mg/kg per day, approximately 45 mg/kg per day, approximately 50 mg/kg per day, approximately 60 mg/kg per day, approximately 70 mg/kg per day, approximately 80 mg/kg per day, approximately 90 mg/kg per day, approximately 100 mg/kg per day, approximately 125 mg/kg per day, approximately 150 mg/kg per day, approximately 175 mg/kg per day, approximately 200 mg/kg per day, approximately 250 mg/kg per day, or approximately 300 mg/kg per day.

In some embodiments, the pharmaceutical combination of the present invention can be administered to provide a systemic effect, such as by intravenous, intraarterial, or intraperitoneal administration, such that the final circulating concentration of the PRMT5 inhibitor compound of formula (I) is from about 0.001 to about 150 micromolar, or greater up to 200 micromolar, 300 micromolar, 400 micromolar, 500 micromolar, 600 micromolar, 700 micromolar, 800 micromolar, 900 micromolar, or 1000 micromolar. The final concentration in the bloodstream (all in micromolar) can be 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 100, 110, 120, 130, 140 or 150 micromolar, or any concentration between any two of the numbers mentioned or greater, as described above, and any concentration within the ranges indicated. Those skilled in the art will determine the required dosages for one or more active agents, taking into account the nature of the agent and the principles discussed herein above.

In some embodiments, the present invention provides a method of administering a PRMT5 inhibitor of formula (I) simultaneously, in parallel, sequentially, one after the other, alternately or separately with at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist or an immune checkpoint inhibitor/modulator.

Co-administration of a PRMT5 inhibitor of formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator may provide an improvement in terms of alleviating or ameliorating one or more symptoms, shortening the duration or reducing the extent of the disease, delaying or slowing the progression of the disease, reducing the intensity, temporarily alleviating or stabilization of the disease state, partial or complete remission, increase in life expectancy and/or other favorable therapeutic results. Such treatments can be administered simultaneously or sequentially in any order with a time interval between administrations. A person skilled in the art will easily determine how to administer the means or treatments, simultaneously, alternately or sequentially and possible time intervals between administrations. When PRMT5 inhibitor compounds are administered at different time points from another modulator of cellular activity described herein, they can be administered for, for example, a subset of 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, or 1 month, or any time interval intermediate between any two of these time intervals. Doses can be administered once daily (QD), twice daily (BID), three times daily (TID), four times daily (QID), or in weekly doses, such as four times per week (QIW), twice per week (BIW), once per week (QW). They can also be administered as needed and before bedtime.

In some embodiments, a treatment according to the present invention may comprise administering to a human subject pharmaceutical combinations comprising a PRMT5 inhibitor of Formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator. For example, the pharmaceutical combinations described herein can be used to prepare a medicinal product for use in improving the general health of a subject or reducing one or more side effects in cancer therapy. Those skilled in the art will readily determine the various agents that will provide a beneficial effect when administered, taking into account, for example, the disease being treated. In some embodiments, when a PRMT5 inhibitor compound of Formula (I) is initially co-administered with at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator, then administration of the corresponding compound of Formula (I) may be discontinued or gradually reduced while administration of the at least one targeted agents/modulators of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator, continues.

The administration of a PRMT5 inhibitor of formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist or an immune checkpoint inhibitor/modulator can be performed at any time during the progression of the disease or disorder or before or after the development of the disorder or one or more symptoms of the disease or disorder. In one embodiment, a PRMT5 inhibitor compound of formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator are administered on a daily basis over an extended period to provide assistance in the ongoing symptom control. In another embodiment, a PRMT5 inhibitor compound of formula (I) and at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator are administered on a daily basis over an extended period of time to either prevent or delay the progression of a disease or disorder.

Synergistic/improved combination

The term "synergistic" as used herein refers to a therapeutic combination that is more effective than the sum of the effects of two or more individual agents. Combination therapy may provide "synergism" and be "synergistic," i.e., exhibit an effect when the active ingredients are administered together that is greater than the sum of the effects resulting from administration of the compounds separately. A synergistic effect may be achieved when the active ingredients are: (1) formulated together and administered or delivered simultaneously in a combined unit dosage formulation, or (2) delivered sequentially as separate formulations. When delivered during sequential therapy, a synergistic effect may be achieved when the compounds are administered or delivered sequentially, such as by different injections in separate syringes. In general, during sequential therapy, an effective dose of each active ingredient is administered sequentially, i.e., sequentially over time. In some embodiments, synergism is evidenced by a lower toxicity of the composition compared to the same dose of either component alone, for the same total dose amount. For example, if a PRMT5 inhibitor compound of formula (I) is co-administered with a cellular activity modulator described herein, the toxicity of a composition comprising a 50:50 (w/w) ratio of the PRMT5 inhibitor compound and the cellular activity modulator described herein is less than the toxicity of 100% (w/w) of the PRMT5 inhibitor compound or 100% (w/w) of the cellular activity modulator, where the composition has approximately the same level of efficacy. Without being limited by theory, the present inventors have found that while a PRMT5 inhibitor compound can be metabolized by a first group of metabolic enzymes, at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator can be metabolized by a second group of enzymes, such that the first group of enzymes and the second group of enzymes are not completely the same. The result is that the toxicity of the combination will be less than that of the individual components of the combination, or that the efficacy of the combination will be greater than that of the individual components of the combination. The rationale for combining two or more types of antitumor therapy is the intention not only to reduce toxicity and provide greater safety, but also to increase the efficacy that a single agent offers as a monotherapy. Increased efficacy is one of the benefits of combination therapy. Combinations are designed to target cellular signaling mechanisms so that several oncogenic signaling pathways can be affected simultaneously, ultimately resulting in increased antitumor activity and/or efficacy (inhibition of tumor growth). Analysis of the examples discussed in this document confirms the improved antitumor effect both in vitro and in vivo , providing a clear example of the benefits of combining compound 24 with other modulators of cellular activity.

When measuring in vivo or therapeutic synergism, one measure of synergism is known as the synergism in excess of the maximum response of either agent alone. Synergism in excess of the maximum response of either agent alone occurs when a fixed dose combination is such that it exceeds both of its component doses, in which case it is called synergism in excess of the maximum response of either agent alone.

An effective amount of a compound (or a pharmaceutically acceptable salt thereof) in the combination of the present invention may be understood to mean an amount that is sufficient to prevent or inhibit tumor cell growth or the progression of cancer metastasis. Therapeutic or pharmacological efficacy of dosages and administration regimens may also be characterized as the ability to induce, enhance, maintain or prolong remission in patients suffering from specific tumors.

Manufacturing and stability

In some embodiments, the formulations of the present invention are substantially pure. By substantially pure is meant that the formulations contain less than about 10%, 5%, or 1%, and preferably less than about 0.1%, of any impurity. In some embodiments, the total amount of impurities, including metabolites of the PRMT5 inhibitor compounds of Formula (I), will be no more than 15%. In some embodiments, the total amount of impurities, including metabolites of the PRMT5 inhibitor compounds of Formula (I), will be no more than 12%. In some embodiments, the total amount of impurities, including metabolites of the PRMT5 inhibitor compounds of Formula (I), will be no more than 11%. In other embodiments, the total amount of impurities, including metabolites of the PRMT5 inhibitor compounds of Formula (I), will be no more than 10%.

In some embodiments, the purity of the compositions of the present invention can be measured using a method selected from anion exchange HPLC (AEX-HPLC) or mass spectrometry. Mass spectrometry can include LC/MS or LC/MS/MS. In some embodiments, the method used to measure the amount of an impurity can include both AEX-HPLC and LC/MS.

Sterile compositions containing the compounds which are PRMT5 inhibitors of formula (I) or pharmaceutically acceptable salts thereof according to the present invention are prepared using aseptic processing by dissolving the compound in a carrier medium for formulation. In one embodiment, the formulation can also be sterilized by filtration. Excipients used in the preparation of the formulations according to the present invention are widely used in pharmaceutical products and are manufactured according to pharmacopoeial standards.

Compositions

Compounds related to the PRMT5 inhibitor compound of formula (I) may be present in a pharmaceutical formulation in a substantially isolated form. It will be understood that the product may be mixed with carriers or diluents that do not interfere with the intended purpose of the product and it will still be considered substantially isolated. The product of the present invention may also be in a substantially purified form, in which case it will generally comprise about 80%, 85% or 90%, such as at least about 88%, at least about 90, 95 or 98% or at least about 99% of a compound of formula (I) of one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist or an immune checkpoint inhibitor/modulator.

Pharmaceutically acceptable diluents, carriers and/or excipients include those that are suitable for veterinary use as well as pharmaceutical use in relation to humans. Suitable carriers and/or excipients will be readily determined by a person of ordinary skill in the art, taking into account the nature of the compounds of formula (I) or other modulator of cellular activity. However, by way of example, diluents, carriers and/or excipients include solutions, solvents, dispersion media, retarding agents, polymeric and lipid agents, emulsions and the like. As a further example, suitable liquid carriers, especially for injection solutions, include water, aqueous saline, aqueous dextrose and the like, with isotonic solutions being preferred for intravenous, intraspinal and intracisternal administration, and carrier media such as liposomes are also particularly suitable for the administration of agents.

In addition, if necessary, substances such as wetting or emulsifying agents, stabilizing or pH buffering agents, or preservatives may also be present in the pharmaceutical combinations of the present invention. In some embodiments, the pharmaceutical combinations of the present invention will contain suitable pharmaceutically acceptable buffers such as acetate buffers, citrate buffers, phosphate buffers, borate buffers, and mixtures thereof. In some embodiments, buffers useful in the present invention include boric acid, sodium borate, sodium phosphates, including mono-, di-, and tribasic phosphates such as monobasic sodium phosphate monohydrate and dibasic sodium phosphate heptahydrate, and mixtures thereof. In some embodiments, the preservative may be stabilized chlorine dioxide, cationic polymers, or quaternary ammonium compounds. In some embodiments, the pharmaceutical combinations may also contain humectants, nutrients, thickeners, antioxidants, and the like, such as disodium ethylenediaminetetraacetate, alkali metal hexametaphosphate, citric acid, sodium citrate, sodium metabisulfite, sodium thiosulfate, N-acetylcysteine, butylated hydroxyanisole, butylated hydroxytoluene, polyvinyl alcohol, poloxamers, polyvinylpyrrolidone, hydroxypropyl methylcellulose, hydroxyethyl methylcellulose, and mixtures thereof. In some embodiments, the pharmaceutical compositions of the present invention do not include a preservative.

In some embodiments, the pharmaceutical combinations of the present invention are in the form of a sterile injectable preparation, such as a sterile injectable aqueous or oily suspension. In some embodiments, the suspension is formulated according to the methods using those suitable dispersing or wetting agents and suspending agents that have been described above. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic diluent or solvent acceptable for parenteral administration, such as a solution in 1,3-butanediol obtained as a lyophilized powder. In some embodiments, acceptable carrier media and solvents that are used can include or exclude water, Ringer's solution (including lactated Ringer's solution), Hartmann's solution, Tyrode's solution and isotonic sodium chloride solution. In some embodiments, sterile fixed oils are used as a solvent or suspending medium. For this purpose, any light, non-volatile oil is used, including synthetic mono- or diglycerides. In some embodiments, fatty acids such as oleic acid are used in the preparation of injectable dosage forms.

The compositions of the present invention may comprise any suitable level of the PRMT5 inhibitor compounds of formula (I) or at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist, or an immune checkpoint inhibitor/modulator, taking into account the dosage form and the mode of administration. However, by way of example, the compositions useful in the present invention may contain from about 0.1% to about 99% by weight, preferably from about 1% to about 60% by weight, of the PRMT5 inhibitor compounds of formula (I), depending on the route of administration.

In addition to standard diluents, carriers and/or excipients, the composition according to the present invention may be formulated with one or more additional constituents or in such a way as to provide an increase in the activity or bioavailability of the compounds of formula (I), wherein at least one targeted agent/modulator of cellular activity selected from an EGFR inhibitor, a KRAS inhibitor, a KRAS-G12C inhibitor, a MEK inhibitor, a Bcl-2 inhibitor, a SOS1 inhibitor, a PARP inhibitor, a RAF inhibitor, an ERK inhibitor, a CDK4/6 inhibitor, a MALT1 inhibitor, a BTK inhibitor, a MAT2A inhibitor, a PI3K inhibitor, an AKT inhibitor, an FGFR inhibitor, a PRMT type I inhibitor, a STING agonist or an immune checkpoint inhibitor/modulator contributes to maintaining their integrity or provides an increase in their half-life or their shelf life, provides sustained release after administration to a subject or, for example, provides other desired advantages. For example, sustained release carrier media include macromers, poly(ethylene glycol), hyaluronic acid, poly(vinylpyrrolidone) or hydrogel. As a further example, the compositions may also include preservatives, solubilizing agents, stabilizing agents, wetting agents, emulsifying agents, sweeteners, coloring agents, flavoring agents, coating agents, buffers, and the like. Those skilled in the art to which the present invention pertains will readily identify additional additives that may be necessary for a particular purpose.

The pharmaceutical combinations described in this document can be formulated according to standard procedures such as those found in standard references such as Gennaro AR: Remington: The Science and Practice of Pharmacy, 20th ed., Lippincott, Williams & Wilkins, 2000.

EXAMPLES

The following examples are provided as illustrations and not as limitations of the methods described herein. Various modifications may be made by those skilled in the art.

Example 1

Materials

The compounds of the present invention can be prepared and characterized using the methods described in International PCT Publication No. WO 2019116302 and described herein. As an illustrative example, the synthesis of compound 24 is described below.

3-Chloro-7-(2-((3aS,4R,6aR)-4-(4-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyl-3a, 6a-dihydro-4H-cyclopenta[d][1,3]dioxol-6-yl)ethyl)-5-fluoroquinoline-2-amine

A mixture of 4-chloro-7-((3aS,4R,6aR)-2,2-dimethyl-6-vinyl-3a,6a-dihydro-4H-cyclopenta[d][1,3]dioxol-4-yl)-7H-pyrrolo[2,3-d]pyrimidine (10.01 g, 31.5 mmol) (prepared according to the procedure described in WO 2019116302, page 116, line 2 to page 117, line 19) and 9-BBN (9-borabicyclononane) (180 mL, 90 mmol) was heated at 50 °C for 1.15 h under N ₂ . The reaction mixture was cooled to room temperature and a solution of tripotassium phosphate (23.88 g, 113 mmol) in water (20 mL) was added and stirred for 30 min. A solution of 7-bromo-3-chloro-5-fluoroquinolin-2-amine (6.2 g, 22.50 mmol) in THF (20 mL) was then added followed by dichloro[1,1'-bis(di-tert-butylphosphino)ferrocene]palladium(II) (1.467 g, 2.250 mmol). The resulting mixture was stirred at 50 °C for 3 h. The reaction mixture was quenched with water (100 mL) and extracted with ethyl acetate (EtOAc) (200 mL). The layers were separated and the organic layer was washed with brine (100 mL) and dried over sodium sulfate. The organic layer was filtered and concentrated in vacuo to give the crude compound. This residue was purified using a CombiFlash (R _f 200, Teledyne/Isco) on a Redisep R _f column with a gradient elution (0 to 40%) of ethyl acetate in dichloromethane to give the title compound (6 g, 51.8%) as an off-white solid. ^1H NMR (400 MHz, DMSO-d6) δ 8.64 (s, 1H), 8.18 (d, J = 0.7 Hz, 1H), 7.25 (s, 1H), 7.09–7.01 (m, 2H), 6.96 (s, 2H), 6.45 (d, J = 3.7 Hz, 1H), 5.67 (s, 1H), 5.56–5.52 (m, 1H), 5.35 (d, J = 5.8 Hz, 1H), 4.52 (d, J = 5.7 Hz, 1H), 3.03 (hept., J = 6.8 Hz, 2H), 2.65 (q, J = 11.9, 8.6 Hz, 2H), 1.38 (s, 3H), 1.29 (s, 3H); LCMS m/z = 515.07 (M+).

7-(2-((3aS,4R,6aR)-4-(4-Amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyl-3a,6a-dihydro-4H-cyclopenta[d][1,3]dioxol-6-yl)ethyl)-3-chloro-5-fluoroquinolin-2-amine

To a stirred solution of 3-chloro-7-(2-((3aS,4R,6aR)-4-(4-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyl-3a,6a-dihydro-4H-cyclopenta[d][1,3]dioxol-6-yl)ethyl)-5-fluoroquinolin-2-amine (6 g, 11.66 mmol) in dioxane (60 mL) was added ammonium hydroxide (114 mL, 2916 mmol) at room temperature. The resulting mixture was stirred at 120 °C for 16 h in a steel autoclave. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (EtOAc) (100 mL). The layers were separated, the organic layer was washed with brine (50 mL), dried over sodium sulfate. The organic layer was filtered and concentrated in vacuo to give the crude compound. This residue was purified using a CombiFlash (R _f 200, Teledyne/Isco) on a Redisep R _f column eluting with a gradient (0 to 8%) of methanol in dichloromethane to give the title compound (4.05 g, 70.1%) as an off-white solid. ^1H NMR (400 MHz, DMSO-d6) δ 8.17 (d, J = 0.7 Hz, 1H), 8.05 (s, 1H), 7.26 (s, 1H), 7.05 (dd, J = 11.1, 1.4 Hz, 1H), 6.97 (d, J = 7.0 Hz, 4H), 6.41 (d, J = 3.5 Hz, 1H), 6.35 (d, J = 3.5 Hz, 1H), 5.53 (d, J = 14.5 Hz, 2H), 5.30 (d, J = 5.7 Hz, 1H), 4.37 (d, J = 5.7 Hz, 1H), 3.02 (h, J = 6.8 Hz, 2H), 2.73–2.58 (m, 2H), 1.37 (s, 3H), 1.27 (s, 3H); LCMS m/z = 495.05 (M+).

(1S,2R,5R)-3-(2-(2-Amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 24)

A mixture of 7-(2-((3aS,4R,6aR)-4-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyl-3a,6a-dihydro-4H-cyclopenta[d][1,3]dioxol-6-yl)ethyl)-3-chloro-5-fluoroquinolin-2-amine (4.05 g, 8.18 mmol) and TFA (26.5 mL, 344 mmol) was stirred at 0°C for 15 min under N₂The resulting mixture was stirred at room temperature for 12 h. The reaction mixture was basified with ice-cold sodium bicarbonate and extracted with ethyl acetate (100 mL). The layers were separated, the organic layer was washed with brine (50 mL) and dried over anhydrous Na₂SO₄. The organic layer was filtered and concentrated.in vacuoto give the crude compound. This residue was purified using a CombiFlash (R_f200, Teledyne/Isco) on a Redisep R column_fwith a gradient elution (0 to 10%) of methanol in dichloromethane to give 3.6 g of the title compound. This compound was further purified by preparative reversed-phase HPLC (YMC Triart C18, 50 x 2.0 mm, 1.9 μm (RFLC-175); mobile phase A – water:CH₃CN (9:1, v/v) + 0.05% NH₄OH, mobile phase B – CH₃CN: water (9:1, v/v) +0.05% NH₄OH; injection volume: 0.2 μL, wavelength: 220 nm, run time: 4.5 min, flow rate: 0.6 mL/min). The compound obtained from preparative reversed-phase (RP) HPLC was co-distilled twice with water (12 mL and 5 mL) to give the title compound (2.3 g, 61.8%) as an off-white solid.¹H NMR (400 MHz, DMSO-d6) δ 8.17 (d, J = 0.8 Hz, 1H), 8.03 (s, 1H), 7.22 (s, 1H), 7.01 (dd , J = 11.0, 1.4 Hz, 1H), 6.95 (s, 2H), 6.92 (s, 2H), 6.63 (d, J = 3.5 Hz, 1H), 6.42 (d, J = 3.5 Hz, 1H), 5.50 (t, J = 3.2 Hz, 1H) , 5.45 (t, J = 1.7 Hz, 1H), 4.96 (dd, J = 6.3, 3.0 Hz, 2H), 4.45 (t, J = 5.8 Hz, 1H), 3.97 (q, J = 5.5 Hz, 1H), 3.03–2.87 (m, 2H), 2.56 (t, J = 7.0 Hz, 2H); LCMS m/z = 454.98 (M+).

Example 2. In vitro data

Compound 24 ((1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol) was tested in combination with targeted therapeutics in vitro , either in a cell viability assay assessed by CellTiter-Glo® (CTG) or in a colony forming ability assay (CFA) assessed by crystal violet staining or CCK8 assay, on a variety of cell lines. Combination studies were performed in the cell lines MDA-MB-231 (breast cancer, TNBC), MIA PaCa-2 (pancreatic cancer with a KRAS mutation), OCI-Ly10 (ABC-diffuse large B-cell lymphoma) and OCI-Ly3 (ABC-diffuse large B-cell lymphoma, a mutant form of CARD11). Both OCI-Ly10 and OCI-Ly3 were obtained under a right-to-use grant from University Health Network, Canada.

Analysis with CTG

MDA-MB-231 cells were seeded in growth medium (DMEM + 10% FBS) at a density of 250 cells/well in a white opaque 96-well plate, followed by overnight incubation under standard cell culture conditions. The next day, cells were treated with compound 24 and another targeting agent as shown in the figures. The treatment was repeated on the 5th day and analysis was completed on the 10th day. Cell viability was assessed at the end of treatment using the CellTiter-Glo® reagent according to the manufacturer's instructions. Luminescence was measured on a BioTek Synergy Neo II plate reader. Relative light units (RLU) are directly proportional to the number of actively proliferating cells.

% inhibition = [[(mean RLU for vehicle medium – mean RLU for blank) – (mean RLU for sample – mean RLU for blank)] × 100]/[(mean RLU for vehicle medium – mean RLU for blank)]

Note: Blank solution contains only CTG and media (no cells)

OCI-Ly3 cells were seeded in growth medium (RPMI + 10% FBS) at a density of 2000 cells/well in a white opaque 96-well plate. Treatment was started on the same day and cells were treated for 4 days. Cell viability was assessed at the end of treatment using the CellTiter-Glo® reagent according to the manufacturer's instructions. Luminescence was measured on a BioTek Synergy Neo II plate reader. The effect on cell viability was assessed for compound 24 in combination with a MALT1 inhibitor.

Analysis with CCK-8

OCI-Ly-10 cells were seeded in growth medium (IMDM + 20% FBS) at a density of 2000 cells/well in a clear 96-well plate. They were treated with different concentrations of the test compounds. Treatment was started on the same day and the cells were treated for 7 days. Cell viability was assessed using the CCK-8 kit (Dojindo Laboratories, China) according to the manufacturer's instructions. The plates were read on a colorimeter and the optical density was determined. (Detection at 450 nm; background correction at 650 nm). The effect on cell viability was assessed for compound 24 in combination with the MALT1 inhibitor or in combination with the BTK inhibitor ibrutinib.

% inhibition (subtracting background level) = [(mean OD for vehicle control – mean OD for sample) × 100]/[mean OD for vehicle control]

Colony Forming Ability Analysis

MIA PaCa-2 cells were seeded at a density of 5,000 cells/well in a 6-well plate or 500 cells/well in a 48-well plate and incubated overnight under standard cell culture conditions. The next day, cells were treated with compound 24 and another targeting agent as shown in the figures. After 7 days of drug treatment, medium was removed from each well and the plates were washed with PBS. Colonies were stained with crystal violet solution for 2-5 min. The plate was then gently washed with tap water and air dried. For quantification, 1,000 μl of dye wash solution containing 10% glacial acetic acid was added to each well and the crystal violet was released from the colonies into solution by sitting on a plate shaker for 20-30 min. After dissolution, the optical density of the extracted dye was recorded on a BioTek Synergy Neo II plate reader at 590 nm. Optical density values were directly proportional to colony growth.

% inhibition = [[(mean optical density for carrier medium – mean optical density for blank solution) – (mean optical density for sample – mean optical density for blank solution)] × 100]/[(mean optical density for carrier medium – mean optical density for blank solution)]

Note: The blank solution contains only dye remover solution.

As shown in Figures 1-7 and further discussed in the in vivo data section below, the combination of compound 24 with various targeted agents (e.g., EGFR inhibitors, SOS1 inhibitors, ERK inhibitors, PARP inhibitors, a KRAS-G12C inhibitor, or CDK4/6 inhibitors) demonstrated synergistic or enhanced inhibition of cell viability compared to each agent alone.

Data in vivo

Example 3

Compound 24 was combined with the EGFR inhibitor afatinib in an in vivo efficacy study in a nude mouse model of human pancreatic cancer using the MIA PaCa-2 cell line xenograft. 20 x 10 ⁶ MIA PaCa-2 cells were injected subcutaneously into nude mice in the presence of PBS and Matrigel at a 1:1 ratio. Tumor-bearing mice were randomized once tumors reached a mean volume of approximately 177-178 mm ³ (with tumor volumes ranging from 119-232 mm ³ ). Mice were divided into the following groups (n=9-10/group): vehicle control, compound 24 (1 mg/kg; twice daily), afatinib (12.5 mg/kg; once daily), and compound 24 + afatinib (compound 24; 1 mg/kg; twice daily + afatinib; 12.5 mg/kg; once daily). Individually, compound 24 and afatinib demonstrated % tumor growth inhibition of 48.03 ± 5.75 and 58.68 ± 5.14, respectively, while the combination of compound 24 and afatinib resulted in 84.16 ± 1.88% tumor growth inhibition.

Furthermore, Fig. 8 demonstrates the unexpected in vivo efficacy of compound 24 with the EGFR inhibitor afatinib in MIA PaCa-2 xenografts.

Example 4

Compound 24 was combined with the SOS1 inhibitor BI-3406 in an in vivo efficacy study in a nude mouse model of human pancreatic cancer using the MIA PaCa-2 cell line xenograft. 20 x 10 ⁶ MIA PaCa-2 cells were injected subcutaneously into nude mice in the presence of PBS and Matrigel at a 1:1 ratio. Tumor-bearing mice were randomized once tumors reached a mean volume of approximately 155-159 mm ³ (with tumor volumes ranging from 107-248 mm ³ ). Mice were divided into the following groups (n=7-8/group): vehicle control, compound 24 (1 mg/kg; twice daily), BI-3406 (50 mg/kg; twice daily), and compound 24 + BI-3406 (compound 24; 1 mg/kg; twice daily + BI-3406; 50 mg/kg; twice daily). Individually, compound 24 and BI-3406 exhibited % tumor growth inhibition of 47.88 ±12.12 and 47.97 ±11.91, respectively, on day 15, while the combination of compound 24 and BI-3406 resulted in 85.98 ± 6.79% tumor growth inhibition on day 15.

Moreover, onFig. 9unexpected effectiveness demonstrated in vivocompound 24 with the SOS1 inhibitor BI-3406 in MIA PaCa-2 xenografts.

Example 5

Compound-24 was combined with the ERK inhibitor ulixertinib (BVD-523) in an in vivo efficacy study in a nude mouse model of human pancreatic cancer using the MIA PaCa-2 cell line xenograft.

MIA PaCa-2 tumor fragments were implanted subcutaneously into nude mice. Tumor-bearing mice were randomized once tumors reached a mean volume of approximately 181-183 ^mm3 (tumor volume ranged from 54-314 ^mm3 ). Mice were divided into the following groups (n=8-10/group): vehicle control, compound 24 (1 mg/kg; twice daily), BVD-523 (25 mg/kg; twice daily), and compound 24 + BVD-523 (compound 24; 1 mg/kg; twice daily + BVD-523; 25 mg/kg; twice daily). Individually, compound 24 and BVD-523 demonstrated % tumor growth inhibition of 58.83 ± 9.92 and 37.48 ± 9.58, respectively, whereas the combination of compound 24 and BVD-523 resulted in 88.05 ± 6.77% tumor growth inhibition.

Furthermore, Fig. 10 demonstrates the unexpected in vivo efficacy of compound 24 with the ERK inhibitor ulixertinib (BVD-523) in MIA PaCa-2 xenografts.

It will be understood that the above description corresponds to an illustrative embodiment of the present invention and that the present invention is not limited to the specific forms shown. Modifications may be made with respect to the structure and arrangement of elements.

The inventions described and claimed herein are characterized by many features and embodiments, including, but not limited to, those shown or described or cited in this detailed description of the invention. The inventions described and claimed herein are not intended to include all the features presented, and are not limited to the features or embodiments shown in this detailed description, which is included solely for illustration and not limitation, or by them. One of ordinary skill in the art will readily recognize that many components and parameters can be changed or modified to a certain extent, or substituted for known equivalents, without departing from the scope of the present invention. It should be noted that such modifications and equivalents are included herein as if they were individually set forth. The present invention also includes all steps, features, compositions and compounds referred to or indicated in this description, individually or in combination, and any and all combinations of any two or more of the stated steps or features.

All patents, publications, scientific articles, websites and other documents and materials cited or referenced in this document indicate the level of skill of those skilled in the art to which the present invention pertains, and each such cited document and material is hereby incorporated by reference to the same extent as if it were individually incorporated by reference in its entirety or set forth herein in its entirety. Applicants reserve the right to physically incorporate into this specification any and all materials and information from any such patents, publications, scientific articles, websites, electronically accessible information sources and other cited materials or documents. Reference to any patent applications, patents and publications in this specification does not constitute and should not be taken as an admission or suggestion in any way that they constitute current prior art or form part of the general public knowledge in any country in the world.

The specific methods and compositions described herein are examples of preferred embodiments and are illustrative and not intended to limit the scope of the present invention. Other objects and embodiments will be apparent to those skilled in the art upon review of this description and are within the scope of the claims. It will be apparent to those skilled in the art that various substitutions and modifications can be made to the present invention disclosed herein. The present invention, illustratively described herein, may suitably be practiced in the absence of any element or elements, or limitation or limitations, not specifically disclosed as key herein. Thus, for example, in each instance herein and in the embodiments or examples of the present invention, any of the terms "comprising," "consisting essentially of," and "consisting of" may be substituted for any of the other two terms in the description. In this regard, the terms "comprising," "including," "having," etc. are to be understood broadly and without limitation. The methods and processes illustratively described herein may appropriately be practiced with a different order of steps and are not necessarily limited to the order of steps recited herein or in the claims. Also, as used herein and in the appended claims, the singular forms include plural references unless the context clearly dictates otherwise. In no event shall a patent be interpreted as limited to the specific examples or embodiments or methods specifically disclosed herein. In no event shall a patent be interpreted as limited by any statement made by any examining examiner or any other officer or employee of the Patent and Trademark Office unless such statement is specifically and without qualification or qualification expressly accepted by applicants in a written response. In addition, titles, headings, or the like are provided to enhance the understanding of the reader of this document and are not to be construed as limiting the scope of this document. Any examples of embodiments, implementations or components of the present invention referred to herein are to be construed as non-limiting.

The terms and expressions which have been used are used as terms of description and not for limitation, and there is no intention to use such terms and expressions to exclude any equivalent of the shown and described features or parts thereof, but it should be obvious that various modifications are possible within the scope of the claimed invention. Thus, it will be understood that although this document has been specifically disclosed by means of preferred embodiments and optional features, a person skilled in the art may resort to modifications and variations of the concepts disclosed in this document, and such modifications and variations are considered to be within the scope of the invention as defined by the appended claims.

The present invention has been described in a broad and general sense herein. Each of the more narrowly defined varieties and subgeneric groups falling within the scope of the disclosure specific to a particular group is also part of the present invention. This includes a general description of the present invention with a condition or negative limitation excluding any subject matter from the genus, whether or not the excluded material is specifically listed herein.

Other embodiments are within the scope of the following claims. Furthermore, where features or embodiments of the present invention are described in terms of Markush groups, one skilled in the art will recognize that the present invention is thereby also described in terms of any individual member or subgroup of members of the Markush group.

Claims (5)

1. A pharmaceutical combination for treating cancer in a subject selected from: (a) the compound (1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 24) or a pharmaceutically acceptable salt thereof and afatinib; (b) the compound (1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 24) or a pharmaceutically acceptable salt thereof and BI-3406; (c) the compound (1S,2R,5R)-3-(2-(2-amino-3-chloro-5-fluoroquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol (compound 24) or a pharmaceutically acceptable salt thereof and ulixertinib. 2. The pharmaceutical combination according to claim 1, wherein the cancer is glioblastoma multiforme, prostate cancer, pancreatic cancer, mantle cell lymphoma, non-Hodgkin's lymphomas and diffuse large B-cell lymphoma, acute myeloid leukemia, acute lymphoblastic leukemia, multiple myeloma, non-small cell lung cancer, small cell lung cancer, breast cancer, triple-negative breast cancer, gastric cancer, colorectal cancer, ovarian cancer, bladder cancer, hepatocellular carcinoma, melanoma, sarcoma, oropharyngeal squamous cell carcinoma, chronic myelogenous leukemia, epidermal squamous cell carcinoma, nasopharyngeal carcinoma, neuroblastoma, endometrial carcinoma, head and neck cancer or cervical cancer.