WO2018081342A1

WO2018081342A1 - Lsd1 inhibitors and uses thereof

Info

Publication number: WO2018081342A1
Application number: PCT/US2017/058404
Authority: WO
Inventors: Victor S. Gehling
Original assignee: Constellation Pharmaceuticals Inc
Current assignee: Constellation Pharmaceuticals Inc
Priority date: 2016-10-26
Filing date: 2017-10-26
Publication date: 2018-05-03
Anticipated expiration: 2019-04-26

Abstract

Provided are novel compounds of Formula (I): and pharmaceutically acceptable salts thereof, which are useful for treating a variety of diseases, disorders or conditions, associated with LSDl. Also provided are pharmaceutical compositions comprising the novel compounds of Formula (I), pharmaceutically acceptable salts thereof, and methods for their use in treating one or more diseases, disorders or conditions, associated with LSDl.

Description

LSD1 INHIBITORS AND USES THEREOF

RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisonal Application No. 62/413,164, filed October 26, 2016, the entire contents of which are incorporated herein by reference.

BACKGROUND

[0002] Lysine- specific demethylase (LSD1), also known as lysine (K)-specific demethylase 1A (LSD1), is a protein in humans that in encoded by the KDM1A gene and specifically demethylates mono- or dimethylated histone H3 lysine4 (H3K4) and H3 lysine 9 (H3K9) via a redox process. (Biochimica et Biophysica Acta 1829 (2013) 981-986). LSD1 has been found to possess oncogenic properties in several cancers ranging from prostate (Cancer Res., 66 (2006), pp. 11341-11347) bladder (Mol. Carcinog., 50 (2011), pp. 931-944) neuroblastomas, (Cancer Res., 69 (2009), pp. 2065-2071) lung cancers, (PLoS One, 7 (2012), p. e35065) sarcomas and hepato-carcinomas (Tumour Biol. (2012). LSD1 pharmacological inhibitors have been shown e.g., to treat leukemias (Nat. Med., 18 (2012), pp. 605-611) and also solid tumors (Tumour Biol. (2012)).

SUMMARY

[0003] It has now been found that compounds of the Formula I:

or a pharmaceutically acceptable salt thereof, and compositions comprising compounds of Formula I are effective inhibitors of LSD1. These compounds were not only found to be effective inhibitors, but it was also surprisingly found that the replacement of ethyl for methyl led to approximately a 28- to 30-fold improvement in potency. See Table 1 in the

Exemplification Section below.

BREIF DESCRIPTION OF THE FIGURES

[0004] FIG. 1 depicts the X-ray crystal structure for intermediate (lR,2S)-2-((E)-l- phenylbut-l-en-2-yl)cyclopropan-l -amine (S)-2-hydroxy-2-phenylacetate. DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

/. Definitions

[0005] The disclosed compounds exist in various stereoisomeric forms. Stereoisomers are compounds that differ only in the spatial arrangement of their atoms. Different spatial arrangements in a compound can result from e.g., the orientation of four different substituents around a chiral carbon atom (i.e., a chiral center), the orientation of two or more substituents around a double bond, or the orientation of two or more substituents on a cycloalkyl ring.

[0006] Enantiomers are one type of stereoisomer that can arise from a chiral center or chiral centers. Enantiomers are pairs of stereoisomers whose mirror images are not superimposable, most commonly because they contain an asymmetrically substituted carbon atom or carbon atoms that acts as a chiral center(s). "R" and "S" represent the absolute configuration of substituents around one or more chiral carbon atoms, where each chiral center is assigned the prefix "R" or "S" according to whether the chiral center configuration is right- (clockwise rotation) or left-handed (counter clockwise rotation). If the turn is clockwise or right-handed about a chiral carbon, the designation is "R" for rectus. If the turn is counter clockwise or left-handed about a chiral carbon, the designation is "S" for sinister.

[0007] Enantiomeric purity reflects the degree to which one enantiomer of a compound is predominantly present over the other enantiomer of that compound. It is determined by subtracting the percent composition of the major enantiomer with the percent composition of the minor enantiomer that is present. For example, a racemic mixture has an enantiomeric purity of 0%, while a single completely pure enantiomer has an enantiomeric purity of 100%. A composition with 70% of one enantiomer and 30% of the other has an enantiomeric purity of 40% (70% - 30%).

[0008] Diastereomers are stereoisomers that are not related as object and mirror image and are not enantiomers. Unlike enantiomers which are mirror images of each other and non- superimposable, diastereomers are not mirror images of each other and non-superimposable. Diastereomers have two or more chiral centers.

[0009] Geometric isomers arise when two or more substituents on a double bond or ring can have different spatial orientations with respect to one another due to the presence of the double bond or ring structure. When the orientation of the substituents of a geometric isomer are on opposite sides of the double bond, those substituents are said to be "trans" to one another or denoted by the letter "E." When the orientation of the substituents of a geometric isomer are on the same side of the double bond, those substituents are said to be "cis" to one another or denoted by the letter "Z."

[0010] When the configuration of two or more substituents about a double bond is indicated by structure; by "E" or "Z" designations; by "cis" or "trans"; or by a combination of the foregoing, it is to be understood that the depicted stereochemical purity with respect to that double bond is at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% by weight. Stereochemical purity by weight with respect to a double bond means the percent by weight of the compound in a composition having the indicated stereochemistry about the double bond. For example, in compounds having the Formula I, when the double bond is represented by

it is to be understood that the compound has a

stereochemical purity with respect to the depicted trans (E) stereochemistry about the double bond, i.e., at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% by weight of the compound in a composition contains the represented trans (i.e., E) double bond.

[0011] Geometric isomers can also arise based on the orientation of two or more substituents about a cyclic group. For example, in compounds of Formula I, the orientation of

about the cyclopropyl can give rise to two different cis configurations

(as in ); and two

differ

trans configurations (as in and

). In instances where the stereochemistry about the cyclopropyl is not defined as in,

the structure includes one of the cis or trans isomers free of other cis and trans stereoisomers, or, alternatively, any mixture of cis and trans stereoisomers.

[0012] When the stereochemistry about the cyclopropyl in the compounds of Formula I is indicated by structure only, the structure is meant to depict the relative stereochemistry at one of the chiral centers in the cyclopropyl relative to the stereochemistry at the other chiral center, and not the absolute stereochemistry at either chiral center in the cyclopropyl. For example, when the stereochemistry about the cyclopropyl is depicted by structure only as being trans, the stereochemical purity of the compound with respect to the depicted trans configuration about the cyclopropyl is at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% by weight, i.e., the percent by weight of the compound in the composition having the trans stereochemistry at the cyclopropyl is at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% by weight. For example, a compound represented by the formula:

means that at least at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% by weight of the compound in the composition has the depicted trans configuration about the cyclopropyl; at least at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% by weight of the compound in the

composition contains the other trans configuration as :

; or at least at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% by weight of the compound in the composition is a mixture of the two trans configurations.

[0013] When the absolute stereochemistry of chiral centers in a compound are indicated structurally and by "R" or "S" designations, it is to be understood that the structure means the depicted stereoisomer at a stereochemical purity of at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% by weight, i.e., the percent by weight of the indicated stereoisomer of the compound in the composition. For example, a compound of Formula I represented by the formula:

means at least 85%, at least 90%, at least 95%, at least

97%, at least 98% or at least 99% by weight of the compound of Formula I in the composition contains of the depicted stereoisomer. When the structure being depicted by structure and by "R" or "S" designation is a single enantiomer, the enantiomeric purity is at least 95% (e.g., at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or at least 99.9%).

[0014] When a compound is depicted structurally without indicating the stereochemistry at a chiral center, it is to be understood that the structure includes either configuration at the chiral center or, alternatively, any mixture of configurations at the chiral center

stereoisomers.

[0015] The 1- and 2- positions of the cyclopropyl ring represent the following:

[0016] The compounds described herein may be present in the form of pharmaceutically acceptable salts. For use in medicines, the salts of the compounds of the invention refer to non-toxic "pharmaceutically acceptable salts." Pharmaceutically acceptable salt forms include pharmaceutically acceptable acidic/anionic or basic/cationic salts. Suitable pharmaceutically acceptable acid addition salts of the compounds described herein include e.g., salts of inorganic acids (such as hydrochloric acid, hydrobromic, phosphoric, nitric, and sulfuric acids) and of organic acids (such as, acetic acid, benzenesulfonic, benzoic, methanesulfonic, and p-toluenesulfonic acids).

[0017] The terms "subject" and "patient" may be used interchangeably, and means a mammal in need of treatment, e.g., companion animals (e.g., dogs, cats, and the like), farm animals (e.g., cows, pigs, horses, sheep, goats and the like) and laboratory animals (e.g., rats, mice, guinea pigs and the like). Typically, the subject is a human in need of treatment. 2. Description of Exemplary Compounds

[0018] In a first embodiment, the present disclosure provides a compound of the Formula I:

or a pharmaceutically acceptable salt thereof.

[0019] In a second embodiment, the present disclosure provides a compound of the Formula III:

or a pharmaceutically acceptable salt thereof.

[0020] In a third embodiment, the present disclosure provides a compound of the Formula IV:

or a pharmaceutically acceptable salt thereof.

[0021] In a fourth embodiment, the present disclosure provides a compound of the Formula V:

or a pharmaceutically acceptable salt thereof.

[0022] In a fifth embodiment, the present disclosure provides a compound of the Formula VI:

or a pharmaceutically acceptable salt thereof. In a sixth embodiment, the present disclosure provides a compound of the

or a pharmaceutically acceptable salt thereof.

[0024] In a seventh embodiment, the present disclosure provides a compound of the Formula VIII:

or a pharmaceutically acceptable salt thereof.

[0025] In an eighth embodiment, the present disclosure provides a compound of the Formula IX:

or a pharmaceutically acceptable salt thereof.

[0026] In a ninth embodiment, the compound described in any one of the fifth, sixth, seventh, or eighth embodiments is a single enantiomer having an enantiomeric purity of at least 95% (e.g., at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or at least 99.9%).

3. Formulation and Administration

[0027] In one aspect, provided herein are compositions comprising the compounds described herein, or a pharmaceutically acceptable salt thereof.

[0028] In another aspect, the compositions further comprise a pharmaceutically acceptable carrier, adjuvant, or vehicle. The amount of compound in the composition is such that is effective to measurably modulate LSD1, or a mutant thereof in a biological sample or in a patient.

[0029] In certain aspects, a composition described herein is formulated for administration to a patient in need of such composition. In some aspects, a composition described herein is formulated for oral administration to a patient. [0030] The term "pharmaceutically acceptable carrier, adjuvant, or vehicle" refers to a non-toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants or vehicles that may be used in the compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose- based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.

[0031] Pharmaceutically acceptable compositions described herein may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.

[0032] Pharmaceutically acceptable compositions described herein may also be prepared in injectable form. Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S. P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

[0033] Pharmaceutically acceptable compositions described herein may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs. Topical application for the lower intestinal tract can be effected in a rectal

suppository formulation (see above) or in a suitable enema formulation. Topically- transdermal patches may also be used.

[0034] The amount of a compound described herein that may be combined with the carrier materials to produce a composition in a single dosage form will vary depending upon the host treated and the particular mode of administration. In some embodiments, provided compositions should be formulated so that a dosage of between 0.01 - 100 mg/kg body weight/day of the inhibitor, such as e.g., 0.1 - 100 mg/kg body weight/day, can be

administered to a patient receiving these compositions.

[0035] It should also be understood that a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease being treated. The amount of compound described herein in the composition will also depend upon the particular compound in the composition.

4. Uses of Compounds and Pharmaceutically Acceptable Compositions

[0036] In some embodiments, the compounds and compositions described herein are useful in treating diseases and/or disorders associated with overexpression of LSD 1 and/or expression of a mutant form of LSD1, such as those mutant forms that alter LSD1 substrate activity.

[0037] In some embodiments, the compounds and compositions described herein are useful in treating diseases and/or disorders associated with cellular proliferation. In some embodiments, the compounds and compositions described herein are useful in treating diseases and/or disorders associated with misregulation of cell cycle or DNA repair. In some embodiments, the compounds and compositions described herein are useful in treating cancer. Exemplary types of cancer include breast cancer, prostate cancer, colon cancer, renal cell carcinoma, glioblastoma multiforme cancer, bladder cancer, melanoma, bronchial cancer, lymphoma and liver cancer.

[0038] In some embodiments, the present disclosure provides a method of reducing the activity of LSD 1 in a subject comprising the step of administering a compound described herein, or a composition comprising any of the compounds herein. In some embodiments, the present disclosure provides a method of reducing the activity of wide-type LSD1 in a subject comprising the step of administering a compound described herein, or a composition comprising any of the foregoing. In some embodiments, the present disclosure provides a method of reducing the activity of a mutant form of LSDl in a subject comprising the step of administering a compound described herein, or a composition comprising any of the foregoing.

[0039] In some embodiments, the present disclosure provides a method of treating a disease or condition related to cancer including e.g., tumors such as skin, breast, brain, cervical carcinomas, testicular carcinomas, etc. In one aspect, cancers that may be treated by the compositions and methods described herein include, but are not limited to tumor types such as astrocytic, breast, cervical, colorectal, endometrial, esophageal, gastric, head and neck, hepatocellular, laryngeal, lung, oral, ovarian, prostate and thyroid carcinomas and sarcomas.

[0040] In some embodiments, the present disclosure provides a method of treating a disease or condition selected from one or more of the following, Cardiac: sarcoma

(angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma and teratoma; Lung: bronchogenic carcinoma (squamous cell,

undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar

(bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma; Gastrointestinal: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma,

leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumors, vipoma), small bowel (adenocarcinoma, lymphoma, carcinoid tumors, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large bowel (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma);

Genitourinary tract: kidney (adenocarcinoma, Wilm's tumor (nephroblastoma), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate ( adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma); Liver: hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor chordoma, osteochronfroma

(osteocartilaginous exostoses), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumors; Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma, osteitis deformans), meninges (meningioma, meningio sarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germinoma (pinealoma), glioblastoma multiform, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), spinal cord neurofibroma, meningioma, glioma, sarcoma);

Gynecological: uterus (endometrial carcinoma), cervix (cervical carcinoma, pre-tumor cervical dysplasia), ovaries (ovarian carcinoma (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa-thecal cell tumors, Sertoli-Leydig cell tumors, dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma), fallopian tubes (carcinoma); Hematologic: blood (myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative diseases, multiple myeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma); Skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, moles dysplastic nevi, lipoma, angioma, dermatofibroma, keloids, psoriasis; and Adrenal glands: neuroblastoma.

[0041] In one embodiment, the present disclosure provides a method of treating a disease or condition seleted from CML, T-ALL, neuroblastoma, breast cancer, prostate cancer, herpes simplex virus reactivation, and HIV infection comprising the step of administering to a subject in need thereof a compound described herein, or a pharmaceutically acceptable salt thereof. In one alternative, the disease or condition is selected from CML, T-ALL, and neuroblastoma.

EXEMPLIFICATION

[0042] The representative examples that follow are intended to help illustrate the present disclosure, and are not intended to, nor should they be construed to, limit the scope of what is described. Modifications and further embodiments, in addition to those shown and described herein, will become apparent to those skilled in the art.

[0043] It will further be appreciated that the present disclosure contemplates individual compounds described herein. Where individual compounds exemplified are isolated and/or characterized as a salt, for example, as a trifluoroacetic acid salt, the present disclosure contemplates a free base of the salt, as well as other pharmaceutically acceptable salts of the free base.

[0044] Unless otherwise noted, all solvents, chemicals, and reagents were obtained commercially and used without purification. The 1H NMR spectra were obtained in CDC1₃, i¾-DMSO, CD₃OD, or d₆-acetone at 25°C at 300 MHz on an OXFORD (Varian) with chemical shift (ό, ppm) reported relative to TMS as an internal standard. HPLC-MS chromatograms and spectra were obtained with Shimadzu LC-MS-2020 system. Chiral analysis and purification were obtained with Yilite P270.

[0045] Preparation of (lR,2S)-2-((E)-l-phenylbut-l-en-2-yl)cyclopropan-l-amine (S)-2-hydroxy-2-phenylac

[0046] Step 1:

(E)-2-benzylidenebutanal

[0047] To a solution of benzaldehyde (130.0kg, 1226.4mol) in MeOH (1100.0kg) was added NaOH (aq.) (10%,500.0kg). Then butanal (92.7kg, 1287.5mol) was added dropwise at 0 °C. The reaction mixture was stirred at 10 °C for 10 h. The solvent was removed and the residue was acidified by HC1 (aq., 4N) to pH = 5. The mixture was extracted with EtOAc (500kg+200kg) and washed with brine (100kg*2). The organic layer was concentrated under vacuum. The crude product was distilled under vacuum (85~95°C, 2~10mm hg) to afford (E)- 2-benzylidenebutanal (130.0kg, GC>95%) as a yellowish oil.

[0048] Step 2:

ethyl (E)-4-((E)-benzylidene)hex-2-enoate

[0049] Ethyl 2-(diethoxyphosphoryl)acetate (182.0kg, 812.5mol) was dissolved in THF (2200kg) and cooled to 0 °C. Potassium ie/t-butanolate (105.0kg, 937.5mol) was added in one portion and the reaction mixture was stirred vigorously at 0 °C for 15 min. (E)-2- benzylidenebutanal (130.0 kg, 812.5mol l.Oeq) was added to the reaction mixture and the reaction mixture was stirred at 25 °C for 16 h. The reaction mixture was quenched with H₂0 (320.0kg) and extracted with EtOAc (360.0kg). The organic layer was dried over sodium sulfate and filtered. The solvent was evaporated under reduced pressure to afford crude product as an oil. The oil was dissolved in petroleum ether (560.0kg), filtered through a pad of silica gel (30.0kg), and the filter cake was washed with petroleum ether. The combined filtrate was concentrated afford ethyl (E)-4-((E)-benzylidene)hex-2-enoate as an oil

(170.0kg). This material was used in the next step without further purification. [0050] Step 3:

ethyl (trans)-2-((E)-l-phenylbut-l-en-2-yl)cyclopropane-l-carboxylate

[0051] Potassium ferf-butanolate (103.5kg, 924.1mol) was dissolved in DMSO (1900kg) under an inert atmosophere. The mixture was stirred for 15 min at 25 °C before addition of trimethylulfoxonium iodide (210.0kg,959.0mol). This mixture was stirred for 45 min at 25 °C before addition of ethyl (E)-4-((E)-benzylidene)hex-2-enoate (170.0kg, 738.0mol). The reaction was warmed to 50-55 °C and stirred for 16 h. The reaction was quenched with water (600kg). The mixture was extracted with petroleum ether (800kg). The petroleum ether phase was evaporated under reduced pressure to afford ethyl (tra«5^,)-2-((E)- l-phenylbut- l-en- 2-yl)cyclopropane-l-carboxylate (76.5kg) as brown liquid. This material was used in the next step without further purification

[0052] Step 4:

(trans)-2-((E)-l-phenylbut-l-en-2-yl)cyclopropane-l-carboxylic acid

[0053] Ethyl (iran5)-2-((E)- l-phenylbut- l-en-2-yl)cyclopropane-l-carboxylate (76.5kg kg, 313.1mol) was dissolved in water (320kg) and methanol (360kg). To this solution was added sodium hydroxide (41.0kg, 1025mol) and the solution was stirred for 3 h at 50 °C. The mixture was concentrated to about 300L and acidified to pH = 4 using HC1 (aq., 4N). This solution was extracted with EtOAc (520kg) and the layers separated. The organic layer was washed with brine, dried over sodium sulfate, filtered, and concentrated to afford (trans)-2- ((E)-l-phenylbut- l-en-2-yl)cyclopropane-l-carboxylic acid (62.5kg). This material was used in next step without further purification.

[0054] Step 5:

(trans)-2-((E)-l-phenylbut-l-en-2-yl)cyclopropan-l-amine hydrochloride

[0055] To a solution of (iran5)-2-((E)- l-phenylbut- l-en-2-yl)cyclopropane-l-carboxylic acid (26.0kg, 120.4mol) in toluene (270kg) were added triethylamine (36.3kg, 359mol) and diphenylphosphoryl azide (38.0kg, 138.1mol) at 0 °C . The reaction mixture was stirred at 0-10 °C for 3 h. Water (100kg) was added and the mixture was extracted with EtOAc (80.0kg) and the organic layer was concentrated under vacuum to afford a crude oil. The crude oil was dissolved in petroleum ether (50 kg) .The suspension was filtered through a pad of silica gel and the filter cake was washed with petroleum ether. The combined filtrates were concentrated to afford the crude acyl azide. The crude acyl azide was taken up in toluene (100kg) and the solution was heated to 85°C for 2 h. The organic layer was concentrated to -60L. Potassium trimethylsilanolate (23.0kg, 177.9mol) was added to the toluene mixture at room temperature and stirred for 1.5 h. The reaction was then treated with HC1 (2N aq., 120kg) and stirred. The layers were separated. The organic layer was extracted twice with HC1 (2N aq., 20kg) and the aqueous extracts were combined. The aqueous layer was extracted with MTBE (30kg) once, and then basified to pH = 10-11 with sodium hydroxide (47kg, 30% aqueous). The aqueous layer was then extracted with MTBE (150kg). The organic extracts were washed with brine, dried with sodium sulfate and filtered. To this MTBE solution was added HC1 (23.5kg, 12% in diethyl ether) and a white solid precipitated from solution. The solid was collected to afford (irafts)-2-((E)-l-phenylbut- l-en-2- yl)cyclopropan- 1 -amine hydrochloride (16.5kg, 73.8mol).

[0056] Step 6:

(lR,2S)-2-((E)-l-phenylbut-l-en-2-yl)cyclopropan-l-amine (S)-2-hydroxy-2- phenylacetate

[0057] (irans)-2-((E)- l-phenylbut- l-en-2-yl)cyclopropan-l -amine hydrochloride (16.5kg, 73.8mol) was charged to the reacter and K₃P0₃(46L, 37wt%) was added followed by EtOAc (150kg). The reaction mixture was stirred at 0-10 °C for 3 h. The organic layer was separated and concentrated to dryness to afford an oil. S-Mandelic acid (8.4kg, 55.35mol) and 95% EtOH (165kg) were added to the reactor and the reaction was stirred at room temperature for 72 h (until IPC ee>99%). The solid was collected via filtration to afford (1R,2S or lS,2R)-2- ((E)- l-phenylbut- l-en-2-yl)cyclopropan-l -amine (S)-2-hydroxy-2-phenylacetate (5.8kg, 17.1 mol). LCMS m/z 188 [M+H]⁺. The absolute stereochemistry was determined by X-ray crystallography.

4-(((lR,2S)-2-((E)-l-phenylbut-l-en-2-yl)cyclopropyl)amino)piperidin-2-one

trifluoroacetate

[0058] A round bottom flask charged with (lR,2S)-2-((E)-l-phenylbut-l-en-2- yl)cyclopropan- 1 -amine (S)-2-hydroxy-2-phenylacetate (500 mg, 1.47 mmol), piperidine-2,4- dione (166 mg, 1.47 mmol), and 5 drops of acetic acid. The reaction was stirred at 60°C in methanol for three hours. Imine formation was followed by LCMS. To this mixture was added sodium cyanoborohydride (277 mg, 4.41 mmol) and the reaction mixture was stirred for 3 hours at 60°C. The reaction was quenched with 10% aqueous potassium carbonate and volatiles were removed under reduced pressure. The remaining residue was partitioned between water/brine (1: 1) and extracted twice with ethyl acetate, dried with sodium sulfate, filtered and evaporated under vacuum. The crude was analyzed by LCMS to reveal a mixture of 85% of the desired product. The crude mixture was purified by reverse phase column chromatography using 10 - 50% C¾CN / 0.1% aq. TFA. The pure fractions were collected, frozen and lyophilized over 24 hours to afford 4-(((lR,2S)-2-((E)-l-phenylbut-l-en-2- yl)cyclopropyl)amino)piperidin-2-one trifluoroacetate (128 mg). LCMS (ESI) m/z: 285.1 [M+H]⁺, RT = 1.22 min.

(R or S)-4-(((lR,2S)-2-((E)-l-phenylbut-l-en-2-yl)cyclopropyl)amino)piperidin-2-one trifluoroacetate

[0059] A round bottom flask charged with (lR,2S)-2-((E)-l-phenylbut-l-en-2- yl)cyclopropan- 1 -amine (S)-2-hydroxy-2-phenylacetate (500 mg, 1.47 mmol), piperidine-2,4- dione (166 mg, 1.47 mmol), and 5 drops of acetic acid was stirred 60°C in methanol for three hours. Imine formation was followed by LCMS. To this mixture was added sodium cyanoborohydride (277 mg, 4.41 mmol) and the reaction mixture was stirred for 3 hours at 60°C. The reaction was quenched with 10% aqueous potassium carbonate and volatiles were removed under reduced pressure. The remaining residue was partitioned between water/brine (1: 1) and extracted twice with ethyl acetate, dried with sodium sulfate, filtered and evaporated under vacuum. The crude was analyzed by LCMS to reveal a mixture of 85% of the desired product. The crude mixture was purified by reverse phase column chromatography using 10 - 50% CH₃CN / 0.1% aq. TFA. The pure fractions were collected, frozen and lyophilized over 24 hours to afford the desired product as a mixture of two disastereomers. The diastereomers were separated by chiral reverse phase SFC (Column: ChiralPak AS, 10 x 250 mm, 5.0 μιη) using C0₂ and z-PrOH/10 mM aq. ammonium formate as eluent. LCMS (ESI) m/z: 285.1 [M+H]⁺, RT = 1.22 min (LCMS Method A).

[0060] Isomer 1, Peak 1: ¹H NMR (400 MHz, DMSO-d₆) δ 9.00 (s, 1H), 8.88 (s, 1H), 7.82 (s, 1H), 7.35 (t, J = 7.5 Hz, 2H), 7.30 - 7.16 (m, 3H), 6.25 (s, 1H), 3.73 (s, 1H), 2.96 (s, 1H), 2.76 - 2.61 (m, 1H), 2.42 - 2.14 (m, 5H), 1.99 (s, 1H), 1.84 - 1.69 (m, 1H), 1.32 - 1.18 (m, 2H), 1.18 - 1.08 (m, 3H).

[0061] Isomer 2, Peak 2: 1H NMR (400 MHz, DMSO-d₆) δ 9.01 (s, 2H), 8.86 (s, 2H), 7.82 (s, 2H), 7.35 (t, J = 7.6 Hz, 4H), 7.22 (dd, J = 12.1, 7.2 Hz, 6H), 6.25 (s, 2H), 3.73 (s, 2H), 2.96 (s, 3H), 2.67 (dd, J = 16.8, 6.0 Hz, 4H), 2.36 - 2.11 (m, 9H), 1.97 (s, 2H), 1.77 (s, 2H), 1.33 - 1.17 (m, 5H), 1.13 (t, J = 7.5 Hz, 6H).

[0062] Each of the aforementioned examples can be reproduced to afford the oppostite enantiomer about the cyclopropyl (i.e., 1S,2R) using the procedures described above and (lS,2R)-2-((E)-l-phenylbut-l-en-2-yl)cyclopropan-l-amine (S)-2-hydroxy-2-phenylacetate instead of (lR,2S)-2-((E)-l-phenylbut-l-en-2-yl)cyclopropan-l -amine (S)-2-hydroxy-2- phenylacetate. The structures of these enantiomers are reproduced below.

LSD1-FRET Inhibition Assay

[0063] LSD1 demethylase reactions were carried out in 50 niM HEPES pH 7.4, 100 niM NaCl, 1 niM DTT, 0.01% Tween-20, and 0.1 mg/niL BSA. All enzymatic reactions were performed for 50 minutes at room temperature in a ΙΟ-μί volume. Five microliters of 8 μΜ biotinylated H3K4mel peptide solution was added to each well of a black 384 well, clear- bottom assay plate containing 80 nL compound (final concentration of 0.8% DMSO and 4 μΜ substrate). Reactions were initiated by the addition of a mixture containing 20 nM LSD1 and 80 nM FAD (5 μί). LSD1 and FAD final concentrations were 10 and 40 nM, respectively. Enzyme activity was stopped by the addition of 90 μΐ_^ of high salt buffer consisting of 50 mM HEPES pH 7.4, 500 mM NaCl, 1 mM DTT, 0.01% Tween-20, and 0.1 mg/mL BSA. Ten microliters of the quenched reaction mixtures were transferred to a black 384 well ProxiPlate. Ten microliters of detection mixture was added to the ProxiPlate, Europium-labeled antibody and Streptavidin APC were used at final concentrations of 0.3 nM and 200 nM, respectively (total assay volume of 20 μί). Capture of the product peptide by the anti-H3K4me0 antibody and Streptavidin APC was allowed to proceed for 60 min at room temperature before measuring the TR-FRET signal. Plates were read on a Perkin Elmer En Vision. Percent inhibition was calculated using Max (no inhibitor) and Min (quenched with stop buffer) controls and inhibition curves plotted to determine IC₅₀ values. Results are shown in Table 1.

LSD1 LY96 Quantigene Assay

[0064] MV4-11 cells were cultured at a density of 4 x 10⁴ cells per well in a 96- well plates and treated with various doses inhibitor starting from 10 μΜ up to 0.0005 μΜ for 16h. The LY-96 mRNA induction was quantified using the Quantigene 2.0 system (Affymetrix). The cells were lysed with Lysis Mixture containing Proteinase K. The working reagent for capturing the RNA was prepared according to the steps detailed in "Capturing Target RNA from Cultured Cell or Blood Lysates" in the Quantigene handbook. The subsequent hybridization with LY-96 probe, signal amplification and detection steps were performed as described in the manual. The chemiluminescence was read using Envision (PerkinElmer) and Abase (IDBS software) was used to plot the dose response curves and calculate IC₅₀. Results are shown in Table 1.

Kasumi-1 GI50 Assay

[0065] Cells were plated at 5,000 cells per well in 96 well tissue culture dishes containing tool compounds arrayed in a 10-point dose curve, ranging from 0 to 10 mM with 4-fold dilutions, and split every fourth day at a fixed ratio to re-establish 5,000 cells/well density for DMSO-treated controls. Cell treatments were carried out for a total of 12 days. At each 4- day split, the viable cell numbers were determined using the Cell Titer-Glo luminescent cell viability assay (Promega, Madison, WI USA) using an En Vision® Multilabel Plate Reader (Perkin Elmer, Waltham, MA USA). GraphPad Prism 6 (GraphPad Software, Inc., La Jolla, CA USA) was used for curve fitting and determination of GI₅₀ values. Results are shown in Table 1.

[0066] As shown above, the disclosed compounds are not only effective LSD1 inhibitors, but they displayed a 28- 30-fold improvement in potency when compared to the methyl analogue comparator compound.

Claims

or a pharmaceutically acceptable salt thereof.

2. The compound of Claim 1, wherein the compound is of the Formula

or a pharmaceutically acceptable salt thereof.

3. The compound of Claim 1 or 2, wherein the compound is of the Formula

or a pharmaceutically acceptable salt thereof.

4. The compound of any ompound is of the Formula

or a pharmaceutically acceptable salt thereof.

5. The compound of any compound is of the Formula

or a pharmaceutically acceptable salt thereof.

6. The compound of Claim 1 or 2, wherein the compound is of the Formula

or a pharmaceutically acceptable salt thereof.

7. The compound of any one of Claims 1 to 3 and 6, wherein the compound is of the Formula

or a pharmaceutically acceptable salt thereof.

8. The compound of any one of Claims 1 to 3 and 6, wherein the compound is of the Formula

or a pharmaceutically acceptable salt thereof.

9. The compound of any one of Claims 4 to 8, wherein the compound is a single enantiomer having an enantiomeric purity of at least 95%.

10. A pharmaceutical composition comprising an effective amount of the compound of any one of Claims 1 to 9 or a pharmaceutically acceptable salt thereof; and a

pharmaceutically acceptable carrier.

11. A composition comprising the compound of any one of Claims 1 to 8, or a pharmaceutically acceptable salt thereof.

12. The composition of Claim 11, wherein the compound is a single enantiomer having an enantiomeric purity of at least 95%.

13. A method of treating a disease or condition selected from CML, T-ALL, neuroblastoma, breast cancer, prostate cancer, herpes simplex virus reactivation, and HIV infection comprising the step of administering to a subject in need thereof the compound of any one of Claims 1 to 9, or a pharmaceutically acceptable salt thereof, or the composition of any one of Claims 10 to 12.

14. The method of Claim 13, wherein the disease or condition is selected from CML, T- ALL, and neuroblastoma.