Telomerase-inhibiting compounds and isolated enantiomers for treating cancer

Abstract

Provided herein are compounds and isolated enantiomers useful for inhibiting telomerase, pharmaceutically acceptable salts thereof, and pharmaceutical compositions comprising at least one such compound, isolated enantiomer, or pharmaceutically acceptable salt thereof. Also described is the use of such compounds, isolated enantiomers, or pharmaceutically acceptable salts thereof, and the use of pharmaceutical compositions comprising at least one such compound, isolated enantiomer, or pharmaceutically acceptable salt thereof, in methods of treating disorders or conditions such as cancer, where the cancer may be a hematological malignancy or a solid tumor.

Description

Attorney Reference: GERN-202WOTELOMERASE-INHIBITING COMPOUNDS AND ISOLATED ENANTIOMERS FOR TREATING CANCERCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 680,925, filed August 8, 2024, the disclosure of which is incorporated herein by reference in its entirety.BACKGROUND

[0002] Telomeres are protective caps at the ends of chromosomes which maintain genomic integrity. In normal, healthy cells, their gradual, progressive shortening over successive cell divisions leads to eventual cell senescence or apoptosis. A majority of cancer cells maintain telomere length by reactivating telomerase, a ribonucleoprotein that catalyzes the addition of telomeric repeat sequences to chromosome ends, thereby avoiding senescence and continuing to divide. Telomere length and telomerase activity are thus significant factors in cancer initiation and survival. Accordingly, telomerase and its inhibition have long been identified as a promising target for cancer therapeutic agents.

[0003] The genes encoding both the protein and RNA components of human telomerase have been cloned and sequenced. The telomerase holoenzyme is composed of the telomerase protein component (the human form of which is known as human telomerase reverse transcriptase or hTERT) and the RNA component (the human form of which is known as human telomerase RNA or hTR). Although hTERT is usually silenced in almost all somatic cells, it is significantly expressed in ~90% of human cancers.

[0004] Much effort has been spent in the search for telomerase inhibitors. Telomerase inhibitors identified to date include small molecule compounds and oligonucleotides. Given the close connection between telomerase and cell proliferative disorders, such as cancer, compounds that inhibit telomerase activity in proliferative cells and the use of the same to treat proliferative cell disorders such as cancer remain an unmet need.Attorney Reference: GERN-202WOSUMMARY OF THE DISCLOSURE

[0005] Described herein are compounds useful for inhibiting telomerase and pharmaceutically acceptable salts thereof; isolated enantiomers of the compounds useful for inhibiting telomerase and pharmaceutically acceptable salts thereof; and pharmaceutical compositions comprising at least one such compound, isolated enantiomer, or pharmaceutically acceptable salt thereof.

[0006] Also described herein are methods of treating diseases or conditions by administering to a subject in need thereof a therapeutically effective amount of at least one compound useful for inhibiting telomerase or a pharmaceutically acceptable salt thereof; at least one isolated enantiomer of the compound useful for inhibiting telomerase or a pharmaceutically acceptable salt thereof; or a pharmaceutical composition comprising at least one such compound, isolated enantiomer, or pharmaceutically acceptable salt thereof.

[0007] In some aspects, the disease or condition is a cancer, where the cancer may be a hematological malignancy or a solid tumor.

[0008] Also provided herein are methods of treating solid tumors by administering to a subject in need thereof a therapeutically effective amount of at least one compound useful for inhibiting telomerase or a pharmaceutically acceptable salt thereof; at least one isolated enantiomer of the compound useful for inhibiting telomerase or a pharmaceutically acceptable salt thereof; or a pharmaceutical composition comprising at least one such compound, isolated enantiomer, or pharmaceutically acceptable salt thereof.BRIEF DESCRIPTION OF THE DRAWINGS

[0009] FIG. 1 shows plasma concentration and time profile for Compound 1 at dosage of 50 mg / kg, according to embodiments of the present disclosure.

[0010] FIG. 2 shows plasma concentration and time profile for Compound 2 at dosage of 50 mg / kg, according to embodiments of the present disclosure.

[0011] FIG. 3 shows plasma concentration and time profile for Compound 3 at dosage of 50 mg / kg, according to embodiments of the present disclosure.

[0012] FIG. 4 shows plasma concentration and time profile for Compound 4 at dosage of 50 mg / kg, according to embodiments of the present disclosure.Attorney Reference: GERN-202WO

[0013] FIG. 5 shows plasma concentration and time profile for Compound 1 at dosage of 10 mg / kg, according to embodiments of the present disclosure.

[0014] FIG. 6 shows plasma concentration and time profile for Compound 3 at dosage of 10 mg / kg, according to embodiments of the present disclosure.

[0015] FIG. 7 shows tissue concentration of Compound 3 at 24 hours, according to embodiments of the present disclosure.

[0016] FIG. 8 shows plasma concentration and time profile for Compound 5 at dosage of 10 mg / kg, according to embodiments of the present disclosure.

[0017] FIG. 9 shows a study design for an efficacy study for ED50 using MOLM13-LUC tumorbearing mice.

[0018] FIG. 10 shows a study design for an efficacy study using MOLM13-LUC tumor-bearing mice.

[0019] FIG. 11 shows the effect of compounds administered on the bioluminescence signal of LUC-MOLM13 tumor-bearing mice, according to embodiments of the present disclosure.

[0020] FIG. 12 shows the effect of compounds administered on the bioluminescence signal of LUC-MOLM13 tumor-bearing mice, according to embodiments of the present disclosure.

[0021] FIG. 13 shows the effect of compounds administered on average body weights of LUC- MOLM13 tumor-bearing mice, according to embodiments of the present disclosure.

[0022] FIG. 14 shows the effect of compounds administered on the life span of LUC-MOLM13 tumor-bearing mice, according to embodiments of the present disclosure.

[0023] FIG. 15 shows the effect of compounds administered on the average radiance in LUC- MOLM13 tumor-bearing mice, according to embodiments of the present disclosure.

[0024] FIG. 16 shows the effect of compounds administered on the bioluminescence signal of LUC-MOLM13 tumor-bearing mice, according to embodiments of the present disclosure.

[0025] FIG. 17 shows LUC-MOLM13 tumors across groups, according to embodiments of the present disclosure.

[0026] FIG. 18 shows a study design for a 4-day acute toxicity study in BALB / c mice, according to embodiments of the present disclosure.

[0027] FIG. 19 shows the effect of compounds administered on the bioluminescence signal of LUC-MOLM13 tumor-bearing mice, according to embodiments of the present disclosure.

[0028] FIG. 20 shows LUC-MOLM13 tumors across groups, according to embodiments of the present disclosure.Attorney Reference: GERN-202WO

[0029] FIG. 21 shows the effect of compounds administered on the bioluminescence signal of LUC-MOLM13 tumor-bearing mice, according to embodiments of the present disclosure.

[0030] FIG. 22 shows the effect of compounds administered on the bioluminescence signal of LUC-MOLM13 tumor-bearing mice, according to embodiments of the present disclosure.

[0031] FIG. 23 shows the effect of compounds administered on the life span of LUC-MOLM13 tumor-bearing mice, according to embodiments of the present disclosure.

[0032] FIG. 24 shows the effect of compounds administered on average body weights of LUC- MOLM13 tumor-bearing mice, according to embodiments of the present disclosure.

[0033] FIG. 25 shows X-ray single crystal data for Compound 2a, according to embodiments of the present disclosure. The stereochemistry of Compound 2a was established as 'S' based on the X-ray data.

[0034] FIG. 26 shows X-ray single crystal data for Compound 2b, according to embodiments of the present disclosure. The stereochemistry of Compound 2b was established as 'R' based on the X-ray data.

[0035] FIG. 27 shows X-ray single crystal data for Compound 4a, according to embodiments of the present disclosure. The stereochemistry of Compound 4a was established as '45,5 / ?' based on the X-ray data.

[0036] FIG. 28 shows X-ray single crystal data for Compound 4b, according to embodiments of the present disclosure. The stereochemistry of Compound 4b was established as '4 / ?, 55' based on the X-ray data.

[0037] FIG. 29 shows X-ray single crystal data for Compound la, according to embodiments of the present disclosure. The stereochemistry of Compound la was established as ' / ?' based on the X-ray data.DETAILED DESCRIPTION

[0038] The present disclosure is not limited to the embodiments described, as such may vary. Moreover, the terminology used herein is for the purpose of describing embodiments only, and is not intended to be limiting.

[0039] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upperAttorney Reference: GERN-202WO and lower limits of that range is also specifically disclosed. Each smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in that stated range is encompassed within the present disclosure. The upper and lower limits of these smaller ranges may independently be included or excluded in the range, and each range where either, neither or both limits are included in the smaller ranges is also encompassed within the present disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the present disclosure.

[0040] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, some potential and exemplary methods and materials may now be described. Any and all publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. It is understood that the present disclosure supersedes any disclosure of an incorporated publication to the extent there is a contradiction.

[0041] As used herein, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a droplet" includes a plurality of such droplets and reference to "the discrete entity" includes reference to one or more discrete entities, and so forth. It is further noted that the claims may be drafted to exclude any element, e.g., any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as "solely", "only" and the like in connection with the recitation of claim elements, or the use of a "negative" limitation.

[0042] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed. To the extent the definition or usage of any term herein conflicts with a definition or usage of a term in an application or reference incorporated by reference herein, the instant application shall control.

[0043] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other severalAttorney Reference: GERN-202WO embodiments without departing from the scope or spirit of the present disclosure. Any recited method can be carried out in the order of events recited or in any other order which is logically possible.DEFINITIONS

[0044] As used herein, unless otherwise specified, reference to an atom is meant to include all isotopes of that atom. For example, reference to H includes1H,2H (i.e., D or deuterium) and3H (i.e., tritium), and reference to C includes both12C and all other isotopes of carbon (e.g.,13C). Unless specified otherwise, groups include all possible stereoisomers.

[0045] As used herein, "acyl" refers to a group of formula -C(O)R wherein R is alkyl, alkenyl, alkynyl, or substituted versions thereof. For example, an acetyl group has formula -C(O)CH3.

[0046] As used herein, "alkenyl" refers to a branched or linear, non-cyclic hydrocarbonyl group that comprises a carbon-carbon double bond. Exemplary alkenyl groups include ethenyl, n-propenyl, isopropenyl, n-butenyl, isobutenyl, octenyl, decenyl, tetradecenyl, hexadecenyl, eicosenyl, and tetracosenyl.

[0047] As used herein, "alkoxy" refers to a group of formula -O(alkyl). Similar groups can be derived from alkenyl, alkynyl, aryl, heteroaryl, and other groups.

[0048] As used herein, "alkyl" refers to a branched or linear, non-cyclic, saturated hydrocarbon group. Exemplary alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, octyl, decyl, cyclopentyl, and cyclohexyl. In some embodiments, the alkyl group has 1 to 24 carbon atoms, e.g. 1 to 12, 1 to 6, or 1 to 3.

[0049] As used herein, the term "alkylene" refers to the divalent version of an alkyl group, i.e., an alkylene group is a divalent, branched or linear, cyclic, or non-cyclic, saturated hydrocarbon group. Exemplary alkylene groups include diylmethane (-CH2-, which is also known as a methylene group), 1,2-diylethane (-CH2CH2-), and 1,1-diylethane (i.e., a CHCH3 fragment where the first atom has two single bonds to other two different groups).

[0050] As used herein, "alkynyl" refers to a branched or linear, non-cyclic hydrocarbonyl group that comprises a carbon-carbon triple bond. Exemplary alkynyl groups include ethynyl and n- propynyl.Attorney Reference: GERN-202WO

[0051] As used herein, "amino" refers to the group -NRXRYwherein Rxand RYare each independently H or a non-hydrogen substituent. Exemplary non-hydrogen substituents include alkyl groups (e.g., methyl, ethyl, and isopropyl).

[0052] As used herein, "aryl" refers to an aromatic group containing at least one aromatic ring, wherein each of the atoms in the ring are carbon atoms, i.e., none of the ring atoms are heteroatoms (e.g., O, S, N). In some embodiments, the aryl group has a second aromatic ring, e.g., that is fused to the first aromatic ring. Exemplary aryl groups are phenyl, naphthyl, biphenyl, diphenylether, diphenylamine, and benzophenone.

[0053] As used herein, the term "arylene" refers to the divalent version of an aryl group, e.g., 1,4- diyl benzene refers to a C6H4 fragment wherein two hydrogens that are located para to one another are removed and replaced with single bonds to other groups.

[0054] As used herein, the term "azido" refers to the group -N3.

[0055] As used herein, "carbonyl" refers to a diradical group of formula -C(O)-.

[0056] As used herein, "carboxy" is used interchangeably with "carboxyl" and "carboxylate" to refer to the -CO2H group and salts thereof.

[0057] As used herein, "cycloalkyl" refers to a cyclic, saturated hydrocarbon group. Similarly, "cycloalkenyl" refers to a cyclic group having a carbon-carbon double bond, whereas "cycloalkynyl" refers to a cyclic group having a carbon-carbon triple bond.

[0058] As used herein, "cyano" or "nitrile" refers to the group -CN.

[0059] As used herein, "ether" refers to a diradical group of formula -O-. For instance, if the ether group is connected to an alkyl group, then the overall group is an alkoxy group (e.g., -OCH3 or methoxy). If the ether is connected to a carbonyl group, then the overall group is an ester group of formula -OC(O)-.

[0060] As used herein, "halo" and "halogen" refer to chloro, bromo, fluoro, or iodo groups.

[0061] "Heteroaryl" refers to an aromatic group containing at least one aromatic ring, wherein at least one of the atoms in the aromatic ring is a heteroatom (e.g., O, S, N). Exemplary heteroaryl groups include those obtained from removing a hydrogen atom from pyridine, pyrimidine, furan, thiophene, or benzothiophene.

[0062] As used herein, "heterocyclyl" refers to a cyclic group that contains a heteroatom (e.g., O, S, N) as a ring atom and that is not aromatic (i.e., distinguishing heterocyclyl groups from heteroarylAttorney Reference: GERN-202WO groups). Exemplary heterocyclyl groups include piperidinyl, tetrahydrofuranyl, dihydrofuranyl, and thiocanyL

[0063] As used herein, "hydroxy" or "hydroxyl" refers to the group -OH.

[0064] As used herein, "imetelstat" refers to the compound with the Chemical Abstract Services (CAS) number of 868169-64-6.

[0065] As used herein, "imetelstat sodium" refers to the sodium salt of imetelstat.

[0066] As used herein, "isolated enantiomer" refers to an enantiomer that has been isolated from other stereoisomers of the compound.

[0067] As used herein, "nitro" refers to a group of formula -NO?.

[0068] As used herein, "pharmaceutically acceptable salt" refers to those salts of the compounds and its isolated enantiomers described herein that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and the like and are commensurate with a reasonable benefit / risk ratio. For example, pharmaceutically acceptable salts are described in: Berge et al., J. Pharmaceutical Sciences 66:1 -19, 1977 and in Handbook of Pharmaceutical Salts: Properties, Selection, and Use, (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. These salts may be acid addition salts involving inorganic or organic acids. The salts can be prepared in situ during the final isolation and purification of the compounds or isolated enantiomers described herein or separately by reacting the free base group with a suitable acid.

[0069] As used herein, "stereoisomers" refers to compounds of the present disclosure that possess asymmetric carbon atoms (optical centers) or double bonds. The racemates, diastereomers, enantiomers, geometric isomers (i.e., cis / trans isomers) and individual stereoisomers of the compounds of the present disclosure are all intended to be encompassed within the scope of the embodiments.

[0070] As used herein, the terms "subject" and "patient" are used interchangeably and refer to any animal, including mammals, and in at least one embodiment, humans.

[0071] As used herein, the term "substituted" refers to the removal of one or more hydrogens from an atom (e.g., from a C or N atom) and their replacement with a different group. For instance, a hydrogen atom on a phenyl (-CeHs) group can be replaced with a methyl group to form a -C6H4CH3 group. Thus, the -C6H4CH3 group can be considered a substituted aryl group. As another example, two hydrogen atoms from the second carbon of a propyl (-CH2CH2CH3) group can be replaced withAttorney Reference: GERN-202WO an oxygen atom to form a -CH2C(O)CHs group, which can be considered a substituted alkyl group. However, replacement of a hydrogen atom on a propyl (-CH2CH2CH3) group with a methyl group (e.g., giving -C^CHfCHs Hs) is not considered a "substitution" as used herein since the starting group and the ending group are both alkyl groups. However, if the propyl group was substituted with a methoxy group, thereby giving a -CH2CH(OCH3)CH3 group, the overall group can no longer be considered "alkyl", and thus is "substituted alkyl". Thus, in order to be considered a substituent, the replacement group is a different type than the original group. In addition, groups are presumed to be unsubstituted unless described as substituted. For instance, the term "alkyl" and "unsubstituted alkyl" are used interchangeably herein.

[0072] As used herein, the term "substituted versions thereof" refers to both substituted and unsubstituted categories being named. For instance, the recitation of "alkyl, aryl, heteroaryl, halo, nitro and substituted versions thereof" refers to the groups alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, and nitro.

[0073] Exemplary substituents include deuterium (D), alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof.

[0074] In some embodiments, the substitutions can themselves be further substituted with one or more groups. For example, the group -C6H4CH2CH3 can be considered as substituted aryl, i.e., an aryl group substituted with the ethyl, which is an alkyl group. Furthermore, the ethyl group can itself be substituted with a pyridyl group to form -C6H4CH2CH2C5H5N, wherein -C6H4CH2CH2C5H5N can also be considered as a substituted aryl group as the term is used herein. In some embodiments, the substituents are not substituted with any other groups.

[0075] As used herein, a "therapeutically effective amount" of a compound, an isolated enantiomer, or a pharmaceutically acceptable salt thereof refers to a quantity of a specified compound, isolated enantiomer, or pharmaceutically acceptable salt thereof sufficient to achieve a desired effect in a subject (e.g., patient) being treated. For example, this may be the amount of a disclosed compound, isolated enantiomer, or pharmaceutically acceptable salt thereof necessary to prevent, inhibit, reduce or relieve a disease or disorder in a subject. In some embodiments, a therapeutically effective amount of a compound, isolated enantiomer, or pharmaceutically acceptable salt thereof is an amount sufficient to prevent, inhibit, reduce or relieve a disease orAttorney Reference: GERN-202WO disorder in a subject without causing a substantial cytotoxic effect on normal host cells in the subject.

[0076] As used herein, "thioether" refers to the group RX-S-RYwherein Rxand RYare each independently a non-hydrogen substituent. Exemplary non-hydrogen substituents include alkyl groups (e.g., methyl, ethyl, and isopropyl).

[0077] As used herein, "thiol" refers to the group -SH.

[0078] As used herein, "treatment" refers to at least an amelioration of at least one symptom associated with the condition afflicting the patient is achieved, where amelioration is used in a broad sense to refer to at least a reduction in the magnitude of a parameter, e.g. symptom, associated with the condition being treated. As such, "treatment" also includes situations where the pathological condition, or at least one symptom associated therewith, is completely inhibited, e.g., prevented from happening, or stopped, e.g. terminated, such that the patient no longer suffers from the condition, or at least at least one symptom that characterize the condition. Thus treatment includes: (i) prevention, that is, reducing the risk of development of at least one clinical symptom, including causing the at least one clinical symptom not to develop, e.g., preventing disease progression to a harmful state; (ii) inhibition, that is, arresting the development or further development of at least one clinical symptom, e.g., mitigating or completely inhibiting an active disease; and / or (iii) relief, that is, causing the regression of at least one clinical symptom.

[0079] As used herein, "unit dosage form" refers to physically discrete units suitable as unitary dosages for human and animal subjects, each unit containing a predetermined quantity of a compound, isolated enantiomer, or pharmaceutically acceptable salt thereof calculated in an amount sufficient to produce the desired effect in association with a pharmaceutically acceptable diluent, excipient, carrier or vehicle. The specifications for a compound, isolated enantiomer, or pharmaceutically acceptable salt thereof depend on the particular compound, isolated enantiomer, or pharmaceutically acceptable salt thereof employed and the effect to be achieved, and the pharmacodynamics associated with each compound, isolated enantiomer, or pharmaceutically acceptable salt thereof in the subject.Attorney Reference: GERN-202WOCOMPOUNDS USEFUL FOR INHIBITING TELOMERASE

[0080] The present disclosure provides compounds and pharmaceutically acceptable salts thereof useful for inhibition of telomerase and in the treatment of cell proliferative disorders such as cancer. In some embodiments, a compound of the present disclosure is described by Formula (I):or a pharmaceutically acceptable salt thereof, wherein: n is 0 or an integer from 1 to 5;R1and R2are independently selected from H, alkyl, cycloalkyl, heterocyclyl, and substituted versions thereof, wherein one of R1and R2is absent; and each R3is independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof, or provided that there are two adjacent R3groups, the two adjacent R3groups along with the atoms to which they are attached can form a cyclic ring.

[0081] As described above, n is 0 or an integer from 1 to 5. In some embodiments, n is an integer from 1 to 4. In some embodiments, n is an integer from 1 to 3. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.

[0082] As described above, R1and R2are independently selected from H, alkyl, cycloalkyl, heterocyclyl, and substituted versions thereof, wherein one of R1and R2is absent. In some embodiments, R1is selected from H, alkyl, cycloalkyl, heterocyclyl, and substituted versions thereof, wherein R2is absent. In some embodiments, R2is selected from H, alkyl, cycloalkyl, heterocyclyl, and substituted versions thereof, wherein R1is absent. In some embodiments, R1and R2are independently selected from H, alkyl, and substituted versions thereof, wherein one of R1and R2is absent. In some embodiments, R1is selected from H, alkyl, and substituted versions thereof, wherein R2is absent. In some embodiments, R2is selected from H, alkyl, and substituted versions thereof, wherein R1is absent. In some embodiments, R1and R2are independently selected from H and methyl, wherein one of R1and R2is absent. In some embodiments, R1is selected from H andAttorney Reference: GERN-202WO methyl, wherein R2is absent. In some embodiments, R2is selected from H and methyl, wherein R1is absent. In certain embodiments, R1is H, wherein R2is absent. In certain embodiments, R1is alkyl or substituted alkyl, such as Ci-6 alkyl or Ci-6 substituted alkyl, or C1-4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl, wherein R2is absent. In certain embodiments, R1is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl, wherein R2is absent. In certain embodiments, R1is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C3-5 heterocycloalkyl or C3-5 substituted heterocycloalkyl, wherein R2is absent. In certain embodiments, R1is Ce heterocycloalkyl or Ce substituted heterocycloalkyl, wherein R2is absent. In certain embodiments, R1is methyl, wherein R2is absent. In certain embodiments, R1is carboxymethyl, wherein R2is absent. In certain embodiments, R2is H, wherein R1is absent. In certain embodiments, R2is alkyl or substituted alkyl, such as C1-6 alkyl or C1-6 substituted alkyl, or C1-4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl, wherein R1is absent. In certain embodiments, R2is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl, wherein R1is absent. In certain embodiments, R2is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C3-5 heterocycloalkyl or C3-5 substituted heterocycloalkyl, wherein R1is absent. In certain embodiments, R2is C& heterocycloalkyl or Ce substituted heterocycloalkyl, wherein R1is absent. In certain embodiments, R2is methyl, wherein R1is absent. In certain embodiments, R2is hydroxyethyl (e.g., 2-hydroxyethyl), wherein R1is absent. In certain embodiments, R2is carboxymethyl, wherein R1is absent. In certain embodiments, R2is piperidinyl, wherein R1is absent. In certain embodiments, R2is tetrahydropyranyl, wherein R1is absent. In certain embodiments, R2is morpholinyl, wherein R1is absent.

[0083] As described above, each R3is independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof, or provided that there are two adjacent R3groups, the two adjacent R3groups along with the atoms to which they are attached can form a cyclic ring. In certain embodiments, R3is alkyl or substituted alkyl, such as C1-6 alkyl or C1-6Attorney Reference: GERN-202WO substituted alkyl, or C1-4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R3is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R3is alkynyl or substituted alkynyl, such as C2-6 alkynyl or C2-6 substituted alkynyl, or C2-4 alkynyl or C2-4 substituted alkynyl, or C2-3 alkynyl or C2-3 substituted alkynyl. In certain embodiments, R3is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, R3is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C3-5 heterocycloalkyl or C3-5 substituted heterocycloalkyl. In certain embodiments, R3is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a C5 aryl or C5 substituted aryl, or a Cg aryl or C& substituted aryl. In certain embodiments, R3is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or Cs-s substituted heteroaryl, such as a C5 heteroaryl or C5 substituted heteroaryl, ora Cg heteroaryl or Ce substituted heteroaryl. In certain embodiments, R3is acyl. In certain embodiments, R3is alkoxy. In certain embodiments, R3is amino. In certain embodiments, R3is azido. In certain embodiments, R3is carbonyl. In certain embodiments, R3is carboxy. In certain embodiments, R3is cyano. In certain embodiments, R3is ether. In certain embodiments, R1is halo. In certain embodiments, R3is hydroxy. In certain embodiments, R3is nitro. In certain embodiments, two adjacent R3groups along with the atoms to which they are attached can form a cyclic ring. In certain embodiments, R3is lactone (e.g., a cyclic carboxylic ester, such as oxolan-2-one) or substituted versions thereof. In certain embodiments, each R3is halo. In certain embodiments, R3is Cl. In certain embodiments, R3is F. In certain embodiments, R3is Br. In certain embodiments, R3is I.

[0084] In some embodiments, a compound of the present disclosure is a compound of Formula (II):or a pharmaceutically acceptable salt thereof,Attorney Reference: GERN-202WO wherein: n is 0 or an integer from 1 to 4; andA is selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof.

[0085] As described above, n is 0 or an integer from 1 to 4. In some embodiments, n is an integer from 1 to 3. In some embodiments, n is 1 or 2. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.

[0086] In some embodiments, A is R3. In some embodiments, R3is A.

[0087] As described above, A is selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof. In certain embodiments, A is alkyl or substituted alkyl, such as Ci-6 alkyl or Ci-6 substituted alkyl, or C1-4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, A is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, A is alkynyl or substituted alkynyl, such as C2-6 alkynyl or C2-6 substituted alkynyl, or C2-4 alkynyl or C2-4 substituted alkynyl, or C2-3 alkynyl or C2-3 substituted alkynyl. In certain embodiments, A is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, A is heterocycloalkyl or substituted heterocycloalkyl, such as C3- 8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C3-5 heterocycloalkyl or C3-5 substituted heterocycloalkyl. In certain embodiments, A is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a C5 aryl or C5 substituted aryl, or a C& aryl or Cg substituted aryl. In certain embodiments, A is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or Cs-s substituted heteroaryl, such as a C5 heteroaryl or C5 substituted heteroaryl, ora Cg heteroaryl or Cg substituted heteroaryl. In certain embodiments, A is acyl. In certain embodiments, A is alkoxy. In certain embodiments, A is amino. In certain embodiments, A is azido. In certain embodiments, A is carbonyl. In certain embodiments, A is carboxy. In certain embodiments, A is cyano. In certain embodiments, A is ether. In certain embodiments, R1is halo. In certain embodiments, A is hydroxy. In certain embodiments, A is nitro. In certain embodiments, A is lactone (e.g., a cyclic carboxylic ester, such as oxolan-2-one) orAttorney Reference: GERN-202WO substituted versions thereof. In certain embodiments, A is Cl. In certain embodiments, A is F. In certain embodiments, A is Br. In certain embodiments, A is I.

[0088] In certain embodiments,, wherein: each R4, R5and R6is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof; each R7is independently H or alkyl; and each R8and R9is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof.

[0089] In certain embodiments,

[0090] As discussed above, each R4, R5and R6is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof. In certain embodiments, R5and R5are independently selected from H, alkyl and hydroxyl.

[0091] In some embodiments, each R4is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof. In certain embodiments, R4is H. In certain embodiments, R4is alkyl or substituted alkyl, such as Ci-6 alkyl or Ci-6 substituted alkyl, or C1-4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R4is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certainAttorney Reference: GERN-202WO embodiments, R4is alkynyl or substituted alkynyl, such as C2-6 alkynyl or C2-6 substituted alkynyl, or C2-4 alkynyl or C2-4 substituted alkynyl, or C2-3 alkynyl or C2-3 substituted alkynyl. In certain embodiments, R4is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, R4is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C3-5 heterocycloalkyl or C3-5 substituted heterocycloalkyl. In certain embodiments, R4is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a C5 aryl or C5 substituted aryl, or a Cg aryl or Cg substituted aryl. In certain embodiments, R4is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or Cs-s substituted heteroaryl, such as a C5 heteroaryl or C5 substituted heteroaryl, ora Cg heteroaryl or Cg substituted heteroaryl. In certain embodiments, R4is acyl. In certain embodiments, R4is alkoxy. In certain embodiments, R4is amino. In certain embodiments, R4is azido. In certain embodiments, R4is carbonyl. In certain embodiments, R4is carboxy. In certain embodiments, R4is cyano. In certain embodiments, R4is ether. In certain embodiments, R4is halo. In certain embodiments, R4is hydroxy. In certain embodiments, R4is nitro. In certain embodiments, R4is thiol. In certain embodiments, R4is thioether. In certain embodiments, each R4is H.

[0092] In some embodiments, each R5is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof. In certain embodiments, R5is H. In certain embodiments, R5is alkyl or substituted alkyl, such as Ci-g alkyl or Ci- substituted alkyl, or C1-4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R5is alkenyl or substituted alkenyl, such as C2- alkenyl or C2-g substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R5is alkynyl or substituted alkynyl, such as C2-g alkynyl or C2-g substituted alkynyl, or C2-4 alkynyl or C2-4 substituted alkynyl, or C2-3 alkynyl or C2-3 substituted alkynyl. In certain embodiments, R5is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C35 cycloalkyl or C35 substituted cycloalkyl. In certain embodiments, R5is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C38 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C3-5 heterocycloalkyl or C3-5 substituted heterocycloalkyl. In certainAttorney Reference: GERN-202WO embodiments, R5is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a C5 aryl or C5 substituted aryl, or a Cg aryl or C& substituted aryl. In certain embodiments, R5is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or Cs-8 substituted heteroaryl, such as a C5 heteroaryl or C5 substituted heteroaryl, ora Cg heteroaryl or Cg substituted heteroaryl. In certain embodiments, R5is acyl. In certain embodiments, R5is alkoxy. In certain embodiments, R5is amino. In certain embodiments, R5is azido. In certain embodiments, R5is carbonyl. In certain embodiments, R5is carboxy. In certain embodiments, R5is cyano. In certain embodiments, R5is ether. In certain embodiments, R5is halo. In certain embodiments, R5is hydroxy. In certain embodiments, R5is nitro. In certain embodiments, R5is thiol. In certain embodiments, R5is thioether. In certain embodiments, R5is selected from H, alkyl and hydroxyl. In certain embodiments, R5is methyl. In certain embodiments, R5is OH.

[0093] In some embodiments, R6is selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof. In certain embodiments, R6is H. In certain embodiments, R6is alkyl or substituted alkyl, such as C1-6 alkyl or C1-6 substituted alkyl, or Ci- 4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R6is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R6is alkynyl or substituted alkynyl, such as C2-6 alkynyl or C2-6 substituted alkynyl, or C2-4 alkynyl or C2-4 substituted alkynyl, or C2-3 alkynyl or C2-3 substituted alkynyl. In certain embodiments, R6is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, R6is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C3-5 heterocycloalkyl or C3-5 substituted heterocycloalkyl. In certain embodiments, R6is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a C5 aryl or C5 substituted aryl, or a Cg aryl or C substituted aryl. In certain embodiments, R6is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or Cs s substituted heteroaryl, such as a C5 heteroaryl or C5 substituted heteroaryl, or a Cg heteroaryl or Cg substituted heteroaryl. In certain embodiments, R6is acyl. In certain embodiments, R6is alkoxy. In certain embodiments, R6is amino. In certain embodiments, R6is azido. In certain embodiments, R6is carbonyl. In certain embodiments, R6is carboxy. In certainAttorney Reference: GERN-202WO embodiments, R6is cyano. In certain embodiments, R6is ether. In certain embodiments, R6is halo. In certain embodiments, R6is hydroxy. In certain embodiments, R6is nitro. In certain embodiments, R6is thiol. In certain embodiments, R6is thioether. In certain embodiments, R6is selected from H, alkyl and hydroxyl. In certain embodiments, R6is methyl. In certain embodiments, R6is OH.

[0094] In certain embodiments,

[0095] In certain embodiments,

[0096] In certain embodiments,

[0097] In certain embodiments,

[0098] As discussed above, each R7is independently H or alkyl. In certain embodiments, R7is H. In certain embodiments, R7is alkyl, such as Ci-6 alkyl, C1-5 alkyl, or C1-4 alkyl, or C1-3 alkyl. In certain embodiments, R7is methyl.

[0099] As discussed above, each R8and R9is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof.

[0100] In some embodiments, each R8is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof. In certain embodiments, R8is H. In certain embodiments, R8is alkyl or substituted alkyl, such as C1-6 alkyl or C1-6 substituted alkyl, or C1-4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R8is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R8is alkynyl or substituted alkynyl, such as C2-6 alkynyl or C2-6 substituted alkynyl, or C2-4 alkynyl or C2-4 substituted alkynyl, or C2-3 alkynyl or C23 substituted alkynyl. In certain embodiments, R8is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C35 cycloalkyl or C35 substitutedAttorney Reference: GERN-202WO cycloalkyl. In certain embodiments, R8is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C3-5 heterocycloalkyl or C3-5 substituted heterocycloalkyl. In certain embodiments, R8is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a C5 aryl or C5 substituted aryl, or a Cg aryl or Cg substituted aryl. In certain embodiments, R8is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or Cs-s substituted heteroaryl, such as a C5 heteroaryl or C5 substituted heteroaryl, ora Cg heteroaryl or Cg substituted heteroaryl. In certain embodiments, R8is acyl. In certain embodiments, R8is alkoxy. In certain embodiments, R8is amino. In certain embodiments, R8is azido. In certain embodiments, R8is carbonyl. In certain embodiments, R8is carboxy. In certain embodiments, R8is cyano. In certain embodiments, R8is ether. In certain embodiments, R8is halo. In certain embodiments, R8is hydroxy. In certain embodiments, R8is nitro. In certain embodiments, R8is thiol. In certain embodiments, R8is thioether. In certain embodiments, each R8is H.

[0101] In some embodiments, R9is selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof. In certain embodiments, R9is H. In certain embodiments, R9is alkyl or substituted alkyl, such as Ci-g alkyl or C1-6 substituted alkyl, or Ci- 4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R9is alkenyl or substituted alkenyl, such as C2-g alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R9is alkynyl or substituted alkynyl, such as C2-6 alkynyl or C2-6 substituted alkynyl, or C2-4 alkynyl or C2-4 substituted alkynyl, or C2-3 alkynyl or C2-3 substituted alkynyl. In certain embodiments, R9is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a Cs-g cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, R9is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C3-5 heterocycloalkyl or C3-5 substituted heterocycloalkyl. In certain embodiments, R9is aryl or substituted aryl, such as Cs s aryl or Cs s substituted aryl, such as a C5 aryl or C5 substituted aryl, or a Cg aryl or Cg substituted aryl. In certain embodiments, R9is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or Cs s substituted heteroaryl, such as a C5 heteroaryl or C5 substituted heteroaryl, or a C heteroaryl or Cg substituted heteroaryl. In certain embodiments, R9is acyl. InAttorney Reference: GERN-202WO certain embodiments, R9is alkoxy. In certain embodiments, R9is amino. In certain embodiments, R9is azido. In certain embodiments, R9is carbonyl. In certain embodiments, R9is carboxy. In certain embodiments, R9is cyano. In certain embodiments, R9is ether. In certain embodiments, R9is halo. In certain embodiments, R9is hydroxy. In certain embodiments, R9is nitro. In certain embodiments, R9is thiol. In certain embodiments, R9is thioether. In certain embodiments, R9is carboxy or cyano.

[0102] In some embodiments, the compound, or a pharmaceutically acceptable salt thereof, has a structure selected from the following structures:Attorney Reference: GERN-202WO

[0103] In some embodiments, the compound useful for inhibiting telomerase is a compound, or a pharmaceutically acceptable salt thereof, listed in the following Table A:Table AAttorney Reference: GERN-202WOAttorney Reference: GERN-202WOAttorney Reference: GERN-202WOISOLATED ENANTIOMERS

[0104] The present disclosure provides isolated enantiomers of compounds and pharmaceutically acceptable salts thereof useful for inhibition of telomerase and in the treatment of cell proliferative disorders such as cancer. In some embodiments, an isolated enantiomer of the present disclosure is described by Formula (I):or a pharmaceutically acceptable salt thereof, wherein: n is 0 or an integer from 1 to 5;R1and R2are independently selected from H, alkyl, cycloalkyl, heterocyclyl, and substituted versions thereof, wherein one of R1and R2is absent; and each R3is independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof, or provided that there are two adjacent R3groups, the two adjacent R3groups along with the atoms to which they are attached can form a cyclic ring.

[0105] As described above, n is 0 or an integer from 1 to 5. In some embodiments, n is an integer from 1 to 4. In some embodiments, n is an integer from 1 to 3. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.

[0106] As described above, R1and R2are independently selected from H, alkyl, cycloalkyl, heterocyclyl, and substituted versions thereof, wherein one of R1and R2is absent. In some embodiments, R1is selected from H, alkyl, cycloalkyl, heterocyclyl, and substituted versions thereof,Attorney Reference: GERN-202WO wherein R2is absent. In some embodiments, R2is selected from H, alkyl, cycloalkyl, heterocyclyl, and substituted versions thereof, wherein R1is absent. In some embodiments, R1and R2are independently selected from H, alkyl, and substituted versions thereof, wherein one of R1and R2is absent. In some embodiments, R1is selected from H, alkyl, and substituted versions thereof, wherein R2is absent. In some embodiments, R2is selected from H, alkyl, and substituted versions thereof, wherein R1is absent. In some embodiments, R1and R2are independently selected from H and methyl, wherein one of R1and R2is absent. In some embodiments, R1is selected from H and methyl, wherein R2is absent. In some embodiments, R2is selected from H and methyl, wherein R1is absent. In certain embodiments, R1is H, wherein R2is absent. In certain embodiments, R1is alkyl or substituted alkyl, such as Ci-6 alkyl or Ci-6 substituted alkyl, or C1-4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl, wherein R2is absent. In certain embodiments, R1is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl, wherein R2is absent. In certain embodiments, R1is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C3-5 heterocycloalkyl or C3-5 substituted heterocycloalkyl, wherein R2is absent. In certain embodiments, R1is Ce heterocycloalkyl or Ce substituted heterocycloalkyl, wherein R2is absent. In certain embodiments, R1is methyl, wherein R2is absent. In certain embodiments, R1is carboxymethyl, wherein R2is absent. In certain embodiments, R2is H, wherein R1is absent. In certain embodiments, R2is alkyl or substituted alkyl, such as C1-6 alkyl or C1-6 substituted alkyl, or C1-4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl, wherein R1is absent. In certain embodiments, R2is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl, wherein R1is absent. In certain embodiments, R2is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C3-5 heterocycloalkyl or C3-5 substituted heterocycloalkyl, wherein R1is absent. In certain embodiments, R2is Ce heterocycloalkyl or Cs substituted heterocycloalkyl, wherein R1is absent. In certain embodiments, R2is methyl, wherein R1is absent. In certain embodiments, R2is carboxymethyl, wherein R1is absent.

[0107] As described above, each R3is independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether,Attorney Reference: GERN-202WO halo, hydroxy, nitro, and substituted versions thereof, or provided that there are two adjacent R3groups, the two adjacent R3groups along with the atoms to which they are attached can form a cyclic ring. In certain embodiments, R3is alkyl or substituted alkyl, such as Ci-6 alkyl or Ci-6 substituted alkyl, or C1-4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R3is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R3is alkynyl or substituted alkynyl, such as C2-6 alkynyl or C2-6 substituted alkynyl, or C2-4 alkynyl or C2-4 substituted alkynyl, or C2-3 alkynyl or C2-3 substituted alkynyl. In certain embodiments, R3is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, R3is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C3-5 heterocycloalkyl or C3-5 substituted heterocycloalkyl. In certain embodiments, R3is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a C5 aryl or C5 substituted aryl, or a Cg aryl or Cg substituted aryl. In certain embodiments, R3is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or Cs-s substituted heteroaryl, such as a C5 heteroaryl or C5 substituted heteroaryl, ora Cg heteroaryl or Cg substituted heteroaryl. In certain embodiments, R3is acyl. In certain embodiments, R3is alkoxy. In certain embodiments, R3is amino. In certain embodiments, R3is azido. In certain embodiments, R3is carbonyl. In certain embodiments, R3is carboxy. In certain embodiments, R3is cyano. In certain embodiments, R3is ether. In certain embodiments, R1is halo. In certain embodiments, R3is hydroxy. In certain embodiments, R3is nitro. In certain embodiments, two adjacent R3groups along with the atoms to which they are attached can form a cyclic ring. In certain embodiments, R3is lactone (e.g., a cyclic carboxylic ester, such as oxolan-2-one) or substituted versions thereof. In certain embodiments, each R3is halo. In certain embodiments, R3is Cl. In certain embodiments, R3is F. In certain embodiments, R3is Br. In certain embodiments, R3is I.

[0108] In some embodiments, the isolated enantiomer is a compound of Formula (II):Attorney Reference: GERN-202WO(II) or a pharmaceutically acceptable salt thereof, wherein: n is 0 or an integer from 1 to 4; andA is selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof.

[0109] As described above, n is 0 or an integer from 1 to 4. In some embodiments, n is an integer from 1 to 3. In some embodiments, n is 1 or 2. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.

[0110] In certain embodiments, A is R3.

[0111] As described above, A is selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof, or provided that there are two adjacent A groups, the two adjacent A groups along with the atoms to which they are attached can form a cyclic ring. In certain embodiments, A is alkyl or substituted alkyl, such as Ci-6 alkyl or Ci-6 substituted alkyl, or C1-4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, A is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, A is alkynyl or substituted alkynyl, such as C2-6 alkynyl or C2-6 substituted alkynyl, or C2-4 alkynyl or C2-4 substituted alkynyl, or C2-3 alkynyl or C2-3 substituted alkynyl. In certain embodiments, A is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, A is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C3-5 heterocycloalkyl or C3-5 substituted heterocycloalkyl. In certain embodiments, A is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a C5 aryl or C5 substituted aryl, or a Cg aryl or Cg substituted aryl. In certain embodiments, A is heteroaryl or substituted heteroaryl, such as Cs s heteroaryl or C5-8 substituted heteroaryl, such as a C5 heteroaryl or C5 substituted heteroaryl, or a Cg heteroaryl or Cg substituted heteroaryl. In certain embodiments, A is acyl. In certain embodiments, A is alkoxy. In certain embodiments, A is amino. In certain embodiments, A is azido.T1Attorney Reference: GERN-202WOIn certain embodiments, A is carbonyl. In certain embodiments, A is carboxy. In certain embodiments, A is cyano. In certain embodiments, A is ether. In certain embodiments, R1is halo. In certain embodiments, A is hydroxy. In certain embodiments, A is nitro. In certain embodiments, A is lactone (e.g., a cyclic carboxylic ester, such as oxolan-2-one) or substituted versions thereof. In certain embodiments, A is Cl. In certain embodiments, A is F. In certain embodiments, A is Br. In certain embodiments, A is I.each R4, R5and R6is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof; each R7is independently H or alkyl; and each R8and R9is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof.

[0113] In certain embodiments,

[0114] As discussed above, each R4, R5and R6is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof. In certain embodiments, R5and R5are independently selected from H, alkyl and hydroxyl.

[0115] In some embodiments, each R4is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof. In certain embodiments, R4is H. In certain embodiments, R4is alkyl or substituted alkyl, such as Ci-6 alkyl or Ci-6 substitutedAttorney Reference: GERN-202WO alkyl, or C1-4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R4is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R4is alkynyl or substituted alkynyl, such as C2-6 alkynyl or C2-6 substituted alkynyl, or C2-4 alkynyl or C2-4 substituted alkynyl, or C2-3 alkynyl or C2-3 substituted alkynyl. In certain embodiments, R4is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, R4is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C3-5 heterocycloalkyl or C3-5 substituted heterocycloalkyl. In certain embodiments, R4is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a C5 aryl or C5 substituted aryl, or a Cg aryl or C& substituted aryl. In certain embodiments, R4is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or Cs-s substituted heteroaryl, such as a C5 heteroaryl or C5 substituted heteroaryl, ora Cg heteroaryl or Ce substituted heteroaryl. In certain embodiments, R4is acyl. In certain embodiments, R4is alkoxy. In certain embodiments, R4is amino. In certain embodiments, R4is azido. In certain embodiments, R4is carbonyl. In certain embodiments, R4is carboxy. In certain embodiments, R4is cyano. In certain embodiments, R4is ether. In certain embodiments, R4is halo. In certain embodiments, R4is hydroxy. In certain embodiments, R4is nitro. In certain embodiments, R4is thiol. In certain embodiments, R4is thioether. In certain embodiments, each R4is H.

[0116] In some embodiments, each R5is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof. In certain embodiments, R5is H. In certain embodiments, R5is alkyl or substituted alkyl, such as C1-6 alkyl or C1-6 substituted alkyl, or C1-4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R5is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R5is alkynyl or substituted alkynyl, such as C2-6 alkynyl or C2-6 substituted alkynyl, or C2-4 alkynyl or C2-4 substituted alkynyl, or C2-3 alkynyl or C2-3 substituted alkynyl. In certain embodiments, R5is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substitutedAttorney Reference: GERN-202WO cycloalkyl. In certain embodiments, R5is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C3-5 heterocycloalkyl or C3-5 substituted heterocycloalkyl. In certain embodiments, R5is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a C5 aryl or C5 substituted aryl, or a Cg aryl or Cg substituted aryl. In certain embodiments, R5is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or Cs-s substituted heteroaryl, such as a C5 heteroaryl or C5 substituted heteroaryl, ora Cg heteroaryl or Cg substituted heteroaryl. In certain embodiments, R5is acyl. In certain embodiments, R5is alkoxy. In certain embodiments, R5is amino. In certain embodiments, R5is azido. In certain embodiments, R5is carbonyl. In certain embodiments, R5is carboxy. In certain embodiments, R5is cyano. In certain embodiments, R5is ether. In certain embodiments, R5is halo. In certain embodiments, R5is hydroxy. In certain embodiments, R5is nitro. In certain embodiments, R5is thiol. In certain embodiments, R5is thioether. In certain embodiments, R5is selected from H, alkyl and hydroxyl. In certain embodiments, R5is methyl. In certain embodiments, R5is OH.

[0117] In some embodiments, R5is selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof. In certain embodiments, R6is H. In certain embodiments, R6is alkyl or substituted alkyl, such as C1-6 alkyl or C1-6 substituted alkyl, or Ci- 4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R6is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R6is alkynyl or substituted alkynyl, such as C2-6 alkynyl or C2-6 substituted alkynyl, or C2-4 alkynyl or C2-4 substituted alkynyl, or C2-3 alkynyl or C2-3 substituted alkynyl. In certain embodiments, Rsis cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, R6is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C35 heterocycloalkyl or C3 s substituted heterocycloalkyl. In certain embodiments, R6is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a C5 aryl or C5 substituted aryl, or a Cg aryl or Cg substituted aryl. In certain embodiments, R6is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or C5-8 substituted heteroaryl, such as a C5 heteroaryl or C5 substitutedAttorney Reference: GERN-202WO heteroaryl, or a Ce heteroaryl or Ce substituted heteroaryl. In certain embodiments, R6is acyl. In certain embodiments, R6is alkoxy. In certain embodiments, Reis amino. In certain embodiments, R6is azido. In certain embodiments, R6is carbonyl. In certain embodiments, R6is carboxy. In certain embodiments, R6is cyano. In certain embodiments, R5is ether. In certain embodiments, R6is halo. In certain embodiments, R6is hydroxy. In certain embodiments, R6is nitro. In certain embodiments, R6is thiol. In certain embodiments, R6is thioether. In certain embodiments, R6is selected from H, alkyl and hydroxyl. In certain embodiments, R6is methyl. In certain embodiments, R6is OH.

[0118] In certain embodiments,

[0119] In certain embodiments,o CR82y ,

[0120] In certain embodiments, A is R .

[0121] In certain embodiments,

[0122] As discussed above, each R7is independently H or alkyl. In certain embodiments, R7is H. In certain embodiments, R7is alkyl, such as Ci-6 alkyl, C1-5 alkyl, or C1-4 alkyl, or C1-3 alkyl. In certain embodiments, R7is methyl.

[0123] As discussed above, each R8and R9is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof.

[0124] In some embodiments, each R8is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof. In certain embodiments, R8is H. In certain embodiments, R8is alkyl or substituted alkyl, such as C1-6 alkyl or C1-6 substituted alkyl, or C1-4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R8is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R8is alkynyl or substituted alkynyl, such as C2-6 alkynyl or C2-6 substituted alkynyl, orAttorney Reference: GERN-202WOC2-4 alkynyl or C2-4 substituted alkynyl, or C2-3 alkynyl or C2-3 substituted alkynyl. In certain embodiments, R8is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, R8is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C3-5 heterocycloalkyl or C3-5 substituted heterocycloalkyl. In certain embodiments, R8is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a C5 aryl or C5 substituted aryl, or a Cg aryl or C& substituted aryl. In certain embodiments, R8is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or Cs-s substituted heteroaryl, such as a C5 heteroaryl or C5 substituted heteroaryl, ora Cg heteroaryl or Cg substituted heteroaryl. In certain embodiments, R8is acyl. In certain embodiments, R8is alkoxy. In certain embodiments, R8is amino. In certain embodiments, R8is azido. In certain embodiments, R8is carbonyl. In certain embodiments, R8is carboxy. In certain embodiments, R8is cyano. In certain embodiments, R8is ether. In certain embodiments, R8is halo. In certain embodiments, R8is hydroxy. In certain embodiments, R8is nitro. In certain embodiments, R8is thiol. In certain embodiments, R8is thioether. In certain embodiments, each R8is H.

[0125] In some embodiments, R9is selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof. In certain embodiments, R9is H. In certain embodiments, R9is alkyl or substituted alkyl, such as C1-6 alkyl or C1-6 substituted alkyl, or Ci- 4 alkyl or C1-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R9is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R9is alkynyl or substituted alkynyl, such as C2-6 alkynyl or C2-6 substituted alkynyl, or C2-4 alkynyl or C2-4 substituted alkynyl, or C2-3 alkynyl or C2-3 substituted alkynyl. In certain embodiments, R9is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, R9is heterocycloalkyl or substituted heterocycloalkyl, such as C3-8 heterocycloalkyl or C3-8 substituted heterocycloalkyl, such as a C3-6 heterocycloalkyl or C3-6 substituted heterocycloalkyl, or a C35 heterocycloalkyl or C35 substituted heterocycloalkyl. In certain embodiments, R9is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a C5 aryl or C5 substituted aryl, or aAttorney Reference: GERN-202WOCg aryl or Cg substituted aryl. In certain embodiments, R9is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or C5-8 substituted heteroaryl, such as a C5 heteroaryl or C5 substituted heteroaryl, or a C& heteroaryl or Cg substituted heteroaryl. In certain embodiments, R9is acyl. In certain embodiments, R9is alkoxy. In certain embodiments, R9is amino. In certain embodiments, R9is azido. In certain embodiments, R9is carbonyl. In certain embodiments, R9is carboxy. In certain embodiments, R9is cyano. In certain embodiments, R9is ether. In certain embodiments, R9is halo. In certain embodiments, R9is hydroxy. In certain embodiments, R9is nitro. In certain embodiments, R9is thiol. In certain embodiments, R9is thioether. In certain embodiments, R9is carboxy or cyano.

[0126] In some embodiments, the isolated enantiomer, or a pharmaceutically acceptable salt thereof, of the present disclosure has a structure selected from the following:Attorney Reference: GERN-202WO

[0127] In some embodiments, the isolated enantiomer is a compound, or a pharmaceutically acceptable salt thereof, listed in the following Table B:Table BAttorney Reference: GERN-202WOAttorney Reference: GERN-202WO

[0128] In some embodiments, the isolated enantiomer, or a pharmaceutically acceptable salt thereof, of the present disclosure has a structure selected from the following:Attorney Reference: GERN-202WOAttorney Reference: GERN-202WOAttorney Reference: GERN-202WO

[0129] In some embodiments, the isolated enantiomer is a compound, or a pharmaceutically acceptable salt thereof, listed in the following Table C:Table CAttorney Reference: GERN-202WOAttorney Reference: GERN-202WOAttorney Reference: GERN-202WOMETHODS OF ISOLATING ENANTIOMERS

[0130] The isolated enantiomers disclosed herein may be obtained via chiral separation methods or enantiomer-selective synthesis methods. In some embodiments, the specific enantiomer is isolated from a racemic mixture using chiral separation methods. Chiral separation methods include, but are not limited to, chromatographic methods, separation using membranes, enantioselective liquid-liquid extractions, and crystallization (see, e.g., Pinto et al., 2020 Molecules 25:1931). Chromatographic separation methods for isolating an enantiomer include, but are not limited to, simulated moving bed chromatography, gas chromatography, liquid chromatography, supercritical fluid chromatography and counter-current chromatography. Chromatographic methods mayAttorney Reference: GERN-202WO include using a chiral column that interacts differently with the different enantiomers, such that a single enantiomer may be isolated and / or purified. Membrane separation methods involve using diffusion enantioselective membranes and / or adsorption-enantioselective membranes that select for a specific enantiomer. Liquid-liquid extractions take advantage of differing solubilities of enantiomers in various solvents, allowing for isolation of a specific enantiomer in a specific solvent. Crystallization methods involve reacting the racemic mixture with chiral resolving agents to obtain derivatives of the racemic mixture. These derivatives of the racemic mixture may then be separated via crystallization to obtain an isolated derivative of the enantiomer, then the isolated derivative of the enantiomer can be converted back into the isolated enantiomer.

[0131] In some embodiments, the isolated enantiomer is synthesized via a stereoselective synthesis (e.g., asymmetric synthesis). In such methods, the synthesis favors the formation of a specific enantiomer. Stereoselective synthesis methods include, but are not limited to, asymmetric catalysis, chiral auxiliary, chiral pool synthesis and biocatalysis. Asymmetric catalysis methods involve using catalysts (e.g., chiral catalysts) that favor synthesis of a specific enantiomer (see, e.g., Trost, 2003 PNAS 101:5348-5355). In chiral auxiliary methods, a chiral auxiliary (e.g., an organic compound) is coupled to a starting material that then is then followed by a synthesis step that favors a certain stereoisomer. The chiral auxiliary is then removed under conditions that preserve the stereoisomer. Chiral pool synthesis involves reacting a chiral starting material in a way such that chirality is preserved throughout the synthesis process. Biocatalysis involves the use of biological catalysts (e.g., enzymes) to produce an enantioselective product. Enzymes may be naturally occurring or non-naturally occurring. In some embodiments, stereoselective synthesis reactions including, but not limited to, asymmetric a I lylation reactions and asymmetric lactonization reactions may be carried out to synthesize an isolated enantiomer (see, e.g., Lazzarotto et al., 2021 Adv Synth Catal 363:3138-3143, Maji et al., 2023 J Am Chem Soc 124:8788-8793, Fukuzawa et aL, 1997 J Am Chem Soc 119:1482-1483).COMPOSITIONS AND FORMULATIONS

[0132] Provided herein are compositions comprising at least one compound, isolated enantiomer, or a pharmaceutically acceptable salt thereof useful for inhibiting telomerase activity. For example, the at least one compound, isolated enantiomer, or pharmaceutically acceptable saltAttorney Reference: GERN-202WO thereof useful for inhibiting telomerase activity can be a compound, isolated enantiomer, or pharmaceutically acceptable salt thereof of Formula (I) or Formula (II) as described above.

[0133] In some embodiments, the composition comprises at least one compound, isolated enantiomer, or pharmaceutically acceptable salt thereof of Formula (I) as described above.

[0134] In some embodiments, the composition comprises at least one compound, isolated enantiomer, or pharmaceutically acceptable salt thereof of Formula (II) as described above.

[0135] In some embodiments, the composition comprises at least one of Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 11, Compound 12, Compound 13, Compound 14, Compound 15, Compound 16, and Compound 17, or a pharmaceutically acceptable salt thereof.

[0136] In some embodiments, the composition comprises at least one isolated enantiomer of Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 11, Compound 12, Compound 13, Compound 14, Compound 15, and Compound 16, or a pharmaceutically acceptable salt thereof.

[0137] In some embodiments, the composition comprises at least one isolated enantiomer selected from Compound la, Compound lb, Compound 2a, Compound 2b, Compound 3a, Compound 3b, Compound 4a, Compound 4b, Compound 10a, Compound 10b, Compound 12a, Compound 12b, Compound 13a, Compound 13b, Compound 14a, Compound 14b, Compound 15a, Compound 15b, Compound 18, and Compound 19, or a pharmaceutically acceptable salt thereof.

[0138] In certain embodiments, the disclosed compounds, isolated enantiomers, and pharmaceutically acceptable salts thereof are useful for the treatment of a disease or disorder. Accordingly, pharmaceutical compositions comprising at least one disclosed compound, isolated enantiomer, or pharmaceutically acceptable salt thereof are also described herein. For example, the present disclosure provides pharmaceutical compositions that include a therapeutically effective amount of at least one compound or isolated enantiomer of the present disclosure (or a pharmaceutically acceptable salt or solvate or hydrate thereof) and a pharmaceutically acceptable excipient, carrier, and / or diluent.

[0139] In some embodiments, the pharmaceutical composition comprises at least one compound, isolated enantiomer, or pharmaceutically acceptable salt thereof of Formula (I) as described above. Optionally, the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient, carrier, and / or diluent.Attorney Reference: GERN-202WO

[0140] In some embodiments, the pharmaceutical composition comprises at least one compound, isolated enantiomer, or pharmaceutically acceptable salt thereof of Formula (II) as described above. Optionally, the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient, carrier, and / or diluent.

[0141] In some embodiments, the pharmaceutical composition comprises at least one of Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 11, Compound 12, Compound 13, Compound 14, Compound 15, Compound 16, and Compound 17, or a pharmaceutically acceptable salt thereof. Optionally, the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient, carrier, and / or diluent.

[0142] In some embodiments, the pharmaceutical composition comprises at least one isolated enantiomer of Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 11, Compound 12, Compound 13, Compound 14, Compound 15, and Compound 16, or a pharmaceutically acceptable salt thereof. Optionally, the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient, carrier, and / or diluent.

[0143] In some embodiments, the pharmaceutical composition comprises at least one isolated enantiomer selected from Compound la, Compound lb, Compound 2a, Compound 2b, Compound 3a, Compound 3b, Compound 4a, Compound 4b, Compound 10a, Compound 10b, Compound 12a, Compound 12b, Compound 13a, Compound 13b, Compound 14a, Compound 14b, Compound 15a, Compound 15b, Compound 18, and Compound 19, or a pharmaceutically acceptable salt thereof. Optionally, the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient, carrier, and / or diluent.

[0144] The pharmaceutical compositions may be manufactured using any of a variety of processes, including, but not limited to, conventional mixing, dissolving, granulating, drageemaking, levigating, emulsifying, encapsulating, entrapping, lyophilizing, and the like.

[0145] At least one compound, isolated enantiomer, or pharmaceutically acceptable salt thereof of the present disclosure may be administered to a subject using any convenient means capable of resulting in the desired reduction in disease, condition, or symptom. Thus, at least one compound, isolated enantiomer, or pharmaceutically acceptable salt thereof of the present disclosure can be incorporated into a variety of formulations for therapeutic administration. More particularly, atAttorney Reference: GERN-202WO least one compound, isolated enantiomer, or pharmaceutically acceptable salt thereof of the present disclosure can be formulated into pharmaceutical compositions by combination with appropriate pharmaceutically acceptable excipients, carriers or diluents, and may be formulated into preparations in solid, semi-solid, liquid or gaseous forms, such as tablets, microtablets, capsules, pills, pellets, powders, lyophilisates, granules, ointments, creams, gels, foams, solutions, suspensions, emulsions, spray-dried dispersions, syrups, suppositories, injections, inhalants, aerosols, and the like. In some embodiments, at least one compound, isolated enantiomer, or pharmaceutically acceptable salt thereof of the present disclosure may be formulated for oral administration (e.g., as an oral dosage). In some embodiments, at least one compound, isolated enantiomer, or pharmaceutically acceptable salt thereof of the present disclosure may be formulated for intravenous administration. In some embodiments, the composition is an aqueous solution.

[0146] Pharmaceutically acceptable carriers useful for the disclosed methods and compositions may depend on the particular mode of administration being employed. In addition to biologically neutral carriers, pharmaceutical compositions to be administered can optionally contain non-toxic auxiliary substances (e.g., excipients), such as wetting or emulsifying agents, preservatives, pH buffering agents, and the like.

[0147] The disclosed pharmaceutical compositions may be formulated with a pharmaceutically acceptable salt of a disclosed compound or isolated enantiomer thereof.

[0148] The dosage form of a disclosed pharmaceutical composition may be determined by the mode of administration chosen. For example, in addition to injectable fluids, topical or oral dosage forms may be employed. Topical preparations may include eye drops, ointments, sprays and the like. Oral formulations may be liquid (e.g., syrups, solutions or suspensions), or solid (e.g., powders, pills, tablets, or capsules). Formulations for intravenous administration may be used, wherein the formulation comprises a suitable carrier. Such suitable carriers include sterile saline although other aqueous and non-aqueous isotonic sterile solutions and sterile suspensions known to be pharmaceutically acceptable are known to those of ordinary skill in the art. Methods of preparing such dosage forms are known, or will be apparent, to those skilled in the art.

[0149] Certain embodiments of the pharmaceutical compositions disclosed herein comprising at least one compound, isolated enantiomer, or pharmaceutically acceptable salt thereof of the present disclosure may be formulated in a unit dosage form suitable for individual administration ofAttorney Reference: GERN-202WO precise dosages. The amount of active ingredient administered may depend on the subject being treated, the severity of the affliction, and the manner of administration, and is known to those skilled in the art. In certain instances, the formulation to be administered contains a quantity of the at least one compound, isolated enantiomer, or pharmaceutically acceptable salt thereof disclosed herein in an amount effective to achieve the desired effect in the subject being treated.

[0150] Each therapeutic compound, isolated enantiomer, or pharmaceutically acceptable salt thereof can independently be in any dosage form, such as those described herein, and can also be administered in various ways, as described herein.

[0151] A disclosed compound, isolated enantiomer, or pharmaceutically acceptable salt thereof can be administered alone, as the sole active pharmaceutical agent, or in combination with one or more additional compounds, isolated enantiomers, or pharmaceutically acceptable salts thereof of the present disclosure or in conjunction with other agents. When administered as a combination, the therapeutic agents can be formulated as separate compositions that are administered simultaneously or at different times, or the therapeutic agents can be administered together as a single composition combining two or more therapeutic agents. Thus, the pharmaceutical compositions disclosed herein containing at least one compound, isolated enantiomer, or pharmaceutically acceptable salt thereof of the present disclosure optionally include one or more other therapeutic agents. Accordingly, certain embodiments are directed to such pharmaceutical compositions, where the composition further includes a therapeutically effective amount of at least one second therapeutic agent.

[0152] Thus, in such embodiments, the pharmaceutical composition comprises at least one disclosed compound, isolated enantiomer, or pharmaceutically acceptable salt thereof (e.g., of Formula (I), (II), or any other formulae described herein) along with at least one second therapeutic agent. The at least one second therapeutic agent may be a telomerase inhibitor, e.g. imetelstat or imetelstat sodium. Imetelstat is a telomerase inhibitor that has been studied for its ability to inhibit various cancers, such as pancreatic cancer (Burchett et al, PLoS One, 2014, 9(1), e85155, doi:10.1371 / journa I. pone.0085155).METHODS OF TREATMENT

[0153] Also provided are methods of treating a patient for a disease or condition by administering to the patient a therapeutically effective amount of at least one disclosed compound, isolatedAttorney Reference: GERN-202WO enantiomer, or a pharmaceutically effective salt thereof (or a pharmaceutical composition comprising at least one of the foregoing). The present disclosure further describes the use of a therapeutically effective amount of at least one compound, isolated enantiomer, or pharmaceutically acceptable salt thereof (or a pharmaceutical composition comprising at least one of the foregoing) as disclosed herein in the treatment of a disease or condition.

[0154] In some embodiments, the methods of treating a patient for a disease or condition disclosed herein comprise administering to the patient in need thereof a therapeutically effective amount of at least one compound, isolated enantiomer, or pharmaceutically acceptable salt thereof of Formula (I) as described above (or a pharmaceutical composition comprising at least one of the foregoing).

[0155] In some embodiments, the methods of treating a patient for a disease or condition disclosed herein comprise administering to the patient in need thereof a therapeutically effective amount of at least one compound, isolated enantiomer, or pharmaceutically acceptable salt thereof of Formula (II) as described above (or a pharmaceutical composition comprising at least one of the foregoing).

[0156] In some embodiments, the methods of treating a patient for a disease or condition disclosed herein comprise administering to the patient in need thereof a therapeutically effective amount of at least one of Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 11, Compound 12, Compound 13, Compound 14, Compound 15, Compound 16, and Compound 17, or a pharmaceutically acceptable salt thereof (or a pharmaceutical composition comprising at least one of the foregoing).

[0157] In some embodiments, the methods of treating a patient for a disease or condition disclosed herein comprise administering to the patient in need thereof a therapeutically effective amount of at least one isolated enantiomer of Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 11, Compound 12, Compound 13, Compound 14, Compound 15, and Compound 16, or a pharmaceutically acceptable salt thereof (or a pharmaceutical composition comprising at least one of the foregoing).

[0158] In some embodiments, the methods of treating a patient for a disease or condition disclosed herein comprise administering to the patient in need thereof a therapeutically effective amount of at least one isolated enantiomer selected from Compound la, Compound lb, Compound 2a, Compound 2b, Compound 3a, Compound 3b, Compound 4a, Compound 4b, Compound 10a,Attorney Reference: GERN-202WOCompound 10b, Compound 12a, Compound 12b, Compound 13a, Compound 13b, Compound 14a, Compound 14b, Compound 15a, Compound 15b, Compound 18, and Compound 19, or a pharmaceutically acceptable salt thereof (or a pharmaceutical composition comprising at least one of the foregoing).

[0159] In some embodiments, the disease or condition is a telomerase-related disease or condition. Such telomerase-related conditions are characterized by telomerase activity or increased telomerase activity or expression or over-expression of telomerase in a cell that normally does not express or expresses at very low levels telomerase or has little or very low telomerase activity. The method can include administering a therapeutically effective amount of at least one compound, isolated enantiomer, or a pharmaceutically acceptable salt thereof (or a pharmaceutical composition comprising at least one of the foregoing) as described above to the patient. In some embodiments, the condition is Hoyeraal-Hreidarsson syndrome, dyskeratosis congenita, pulmonary fibrosis, aplastic anemia, or liver fibrosis.

[0160] In some embodiments, the disease or condition is a cancer. In some embodiments, the cancer is a hematological malignancy or a solid tumor.

[0161] Cancers that are solid tumors represent approximately 90% of all cancers in adults. Solid tumor cancers include, but are not limited to, cancers of the lung, colon, central nervous system (e.g., brain and spinal cord), skin, ovary, kidney, prostate and breast.

[0162] Hematologic malignancies are forms of cancer that begin in the cells of blood-forming tissue, such as the bone marrow, or in the cells of the immune system. Examples of hematologic malignancies are acute and chronic leukemias, lymphomas, multiple myeloma, myelodysplastic syndromes, and myeloproliferative neoplasms.

[0163] Cancers that are solid tumors are abnormal masses of tissues. They represent approximately 90% of all cancers in adults. Solid tumor cancers include, but are not limited to, cancers of the lung; colon and / or colorectal; central nervous system (e.g., brain and spinal cord), such as glioma, glioblastoma, head and neck squamous cell carcinoma; skin (such as melanoma); ovary; cervix; endometrium; kidney; prostate; bladder; liver; breast; and esophagus. Types of solid tumors include sarcomas and carcinomas. Sarcomas are cancers that begin in the bones and soft tissues. Such soft tissues include muscle, fat, blood vessels, nerves, tendons and linings of the joints. Carcinomas are cancers that begin in epithelial tissues. Epithelial tissues line most organs and internal passageways ofAttorney Reference: GERN-202WO the body. Non-limiting examples of carcinomas include adrenocortical carcinoma, hepatocellular carcinoma, and stomach adenocarcinoma.

[0164] In some embodiments, the cancer is selected from acute and chronic leukemias, lymphomas, multiple myeloma and myelodysplastic syndromes, myeloproliferative neoplasms (MPNs), essential thrombocythemia (ET), polycythemia vera (PV), Chronic Myelogenous Leukemia (CML), myelofibrosis (MF), acute myelogenous leukemia (AML), myelodysplastic syndromes (MDS), lung cancers, colon cancers, central nervous system (CNS) cancers, skin cancers, ovarian cancers, renal cancers, prostate cancers and breast cancers.

[0165] Leukemia

[0166] In some embodiments, the cancer is a leukemia. Leukemia is a group of blood cancers. The four main types of leukemia are acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL) and chronic myeloid leukemia (CML). AML is a cancer of the myeloid line of blood cells. AML is the most common acute leukemia affecting adults. Patients with AML have a rapid growth of abnormal white blood cells that accumulate in the bone marrow and interfere with the production of normal blood cells. Replacement of normal bone marrow with leukemic cells causes a drop in red blood cells, platelets, and normal white blood cells. The symptoms of AML include fatigue, shortness of breath, easy bruising and bleeding, and increased risk of infection. However, there are many other less common types of leukemia, as well. Lymphomas are blood cancers that affect lymphocytes. Multiple myeloma is cancer of plasma cells, wherein the plasma cells produce abnormal antibodies. Myeloproliferative neoplasm is a group of cancers where the bone marrow makes too many blood cells (e.g., red blood cells, white blood cells or platelets).

[0167] Essential Thrombocythemia

[0168] In some embodiments, the cancer is Essential Thrombocythemia. Circulating blood platelets are anucleate, although they retain small amounts of megakaryocyte-derived mRNAs and a fully functional protein biosynthetic capacity (Gnatenko et al., Blood 101, 2285-2293 (2003)). Essential Thrombocythemia (ET) is a myeloproliferative disorder subtype, characterized by increased neoplastic proliferation of megakaryocytes, elevated numbers of circulating platelets, and considerable thrombohemorrhagic events, not infrequently neurological (Nimer, Blood 93, 415-416 (1999)).Attorney Reference: GERN-202WO

[0169] Polycythemia Vera

[0170] In some embodiments, the cancer is Polycythemia Vera. Patients with Polycythemia Vera (PV) have marked increases of red blood cell production. Treatment is directed at reducing the excessive numbers of red blood cells. PV can develop a phase late in their course that resembles primary myelofibrosis with cytopenias and marrow hypoplasia and fibrosis.

[0171] Myelofibrosis

[0172] In some embodiments, the cancer is myelofibrosis. Myelofibrosis or MF, or primary myelofibrosis, is a myeloproliferative neoplasm in the same spectrum of diseases as ET. Occasionally ET evolves into MF.

[0173] Myelodysplastic syndrome

[0174] In some embodiments, the cancer is myelodysplastic syndrome. Myelodysplastic syndromes (MDS) represent a rare heterogenous group of hemopoietic clonal disorders that are characterized by ineffective hemopoiesis resulting in anemia and other cytopenias and are serious and life threatening with a high risk of leukemic transformation. Myelodysplastic syndromes (MDS) include diseases, such as refractory anemia, refractory anemia with excess blasts, refractory cytopenia with multilineage dysplasia, refractory cytopenia with unilineage dysplasia, and chronic myelomonocytic leukemia. The immature blood stem cells (blasts) do not become healthy red blood cells, white blood cells or platelets. The blasts die in the bone marrow or soon after they travel to the blood. This leaves less room for healthy white cells, red cells and / or platelets to form in the bone marrow. The myelodysplastic syndromes (MDS) are a collection of hematological medical conditions that involve ineffective production of the myeloid class of blood cells. Patients with MDS often develop severe anemia and require frequent blood transfusions. Bleeding and risk of infections also occur due to low or dysfunctional platelets and neutrophils, respectively. In some cases, the disease worsens and the patient develops cytopenias (low blood counts) caused by progressive bone marrow failure. In some cases, the disease transforms into acute myelogenous leukemia (AML).

[0175] Lung cancers

[0176] In some embodiments, the cancer is lung cancer. Lung cancers include small cell lung cancers (SCLC) and non-small cell lung cancers (NSCLC). The majority of lung cancers are NSCLCs. NSCLCs include adenocarcinomas, squamous-cell carcinomas, and large-cell carcinomas. Nearly 40% of lung cancers are adenocarcinomas and about 30% of lung cancers are squamous-cell carcinomas.Attorney Reference: GERN-202WO

[0177] Bladder cancers

[0178] In some embodiments, the cancer is bladder cancer. Bladder cancers refer to several types of cancer that arise from the tissue of the urinary bladder. Smoking greatly increases the risk of bladder cancer. The most common type of bladder cancer is transitional cell carcinoma. Other types of bladder cancer include, but are not limited to, squamous cell carcinoma and adenocarcinoma.

[0179] Liver cancers

[0180] In some embodiments, the cancer is liver cancer. Liver cancer, also known as hepatic cancer, is cancer of the liver tissue. Primary hepatic cancer refers to cancer that originates in the liver. Secondary hepatic cancer, also known as liver metastasis, is cancer that spreads from elsewhere in the body to the liver. Secondary hepatic cancer is more common than primary hepatic cancer. Hepatocellular carcinoma (HCC) is the most common type of cancer. Other types of liver cancer include, but are not limited to, intrahepatic cholangiocarcinoma, angiosarcoma, hemangiosarcoma, and hepatoblastoma. Viral infection with hepatitis C virus or hepatitis B virus greatly increases the risk of liver cancer (e.g., HCC).

[0181] Colon cancers

[0182] In some embodiments, the cancer is colon cancer. Colon cancer, which is also known as bowel cancer or colorectal cancer, is the development of a cancer tumor within the colon or rectum of the large intestine. The three main types of colon cancer include sporadic, hereditary and colitis- associated colon cancer. The risk of colon cancer is increased with other diseases including, but not limited to, inflammatory bowel disease, Crohn's disease and ulcerative colitis.

[0183] Adrenal cancers

[0184] In some embodiments, the cancer is adrenal. Adrenal cancer, such as adrenocortical carcinoma (ACC) is a cancer originating from the cortex of the adrenal gland. ACC is a rare cancer that is possibly linked to genetic mutations.

[0185] Esophageal cancers

[0186] In some embodiments, the cancer is esophageal cancer. Esophageal cancers originate from the esophagus. The two main types of esophageal cancer are esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EAC). ESCC originates from the epithelial cells that line the esophagus. EAC originates from the glandular cells present in the lower portion of the esophagus.Attorney Reference: GERN-202WO

[0187] Stomach cancers

[0188] In some embodiments, the cancer is stomach cancer. Stomach cancers, also known as gastric cancers, develop from the lining of the stomach. The majority of stomach cancers are gastric carcinomas, including gastric adenocarcinoma. The most common cause of stomach cancer is bacterial infection by Helicobacter pylori.

[0189] Central nervous system (CNS) cancers

[0190] In some embodiments, the cancer is central nervous system cancer. CNS cancers include cancers of the brain and the spinal cord. CNS tumors are typically discovered when patients develop symptoms. CNS cancers include, but are not limited to, gliomas, glioblastomas, astrocytomas, meningiomas, oligodendrogliomas, ependymomas, mixed gliomas, mixed glial and neuronal tumors (e.g., ganglioglioma, pleomorphic xanthoastrocytoma, and dysembryoplastic neuroepithelial tumor), and primitive neuroectodermal tumors (e.g., medulloblastoma, pineoblastomas, neuroblastomas).

[0191] Skin cancers

[0192] In some embodiments, the cancer is skin cancer. Skin cancers include, but are not limited to, basal-cell skin cancer, squamous-cell skin cancer, such as head and neck squamous cell carcinoma, and melanoma. Of these, melanomas are the most aggressive type of skin cancer.

[0193] Ovarian cancers, cervical cancers and endometrial cancers

[0194] In some embodiments, the cancer is ovarian cancer. Ovarian cancers may originate from the ovary or from nearby structures such as the fallopian tubes. Ovarian cancer can be related to genetic risk factors including, but not limited to, mutations of the BRCA1 or BRCA2 genes, hereditary nonpolyposis colon cancer and Peutz-Jeghers syndrome. More than 95% of ovarian cancers are epithelial ovarian carcinomas.

[0195] In some embodiments, the cancer is cervical cancer. Cervical cancers generally originate from the cervix or any layer of the wall of the cervix. Human papillomavirus infection (HPV) causes more than 90% of cases of cervical cancer. Approximately 90% of cervical cancers are squamous cell carcinomas.

[0196] In some embodiments, the cancer is endometrial cancer. Endometrial cancers originate from the endometrium, which is the lining of the uterus. 80% of endometrial cancers are endometrioid carcinomas.Attorney Reference: GERN-202WO

[0197] Renal cancers

[0198] In some embodiments, the cancer is renal cancer. Renal cancers are cancers of the kidney. Renal cancers include, but are not limited to, renal cell cancer (RCC), transitional cell cancer (TCC) and Wilms' tumor. Nearly 80% of renal cancers are RCCs.

[0199] Prostate cancers

[0200] In some embodiments, the cancer is prosate cancer. Prostate cancers include, but are not limited to adenocarcinomas, small cell carcinomas, neuroendocrine tumors, transitional cell carcinomas and sarcomas. Nearly all prostate cancers are adenocarcinomas.

[0201] Breast cancers

[0202] In some embodiments, the cancer is breast cancer. Breast cancers generally begin in the lobules or ducts of the breasts. Invasive ductal carcinoma (I DC) is a breast cancer that begins in the milk ducts. IDC is the most common type of breast cancer with approximately 80% of all breast cancers being IDC. Invasive lobular carcinoma (ILC) is a breast cancer that begins in the lobules, which are the glands that produce milk.

[0203] In some embodiments, a disclosed compound, isolated enantiomer, or pharmaceutically acceptable salt thereof is administered in a therapeutically effective amount to a patient to treat a disease or condition. In some instances, the administration of the therapeutically effect amount can inhibit the telomerase in cancer cells of the patient, thereby inhibiting the cancer. In some instances, the administration may inhibit the cancer by another biological mechanism. In some cases, a disclosed compound, isolated enantiomer, or pharmaceutically acceptable salt thereof can be administered to the patient, and the method can further comprise administering to the patient an additional compound or at least one second therapeutic agent, e.g., a telomerase inhibitor such as imetelstat or imetelstat sodium.

[0204] The disclosed compounds, isolated enantiomers, and pharmaceutically acceptable salts thereof are useful for treating a disease, disorder or condition in a subject. The route of administration may be selected according to a variety of factors including, but not limited to, the condition to be treated, the formulation and / or device used, the subject to be treated, and the like. Routes of administration useful in the disclosed methods include, but are not limited to, oral and parenteral routes, such as intravenous (iv), intraperitoneal (ip), rectal, topical, ophthalmic, nasal, otic, intrathecal, and transdermal. Formulations for these dosage forms are described herein.Attorney Reference: GERN-202WO

[0205] A therapeutically effective amount of a disclosed compound, isolated enantiomer, or pharmaceutically effective salt thereof may depend, at least, on the particular method of use, the subject being treated, the severity of the affliction, and the manner of administration of the therapeutic composition.

[0206] Therapeutically effective doses of a subject compound or isolated enantiomer or pharmaceutical composition can be determined by one of skill in the art. For example, in some instances, a therapeutically effective dose of a compound or isolated enantiomer or pharmaceutical composition is administered with a goal of achieving local (e.g., tissue) or plasma concentrations that are at least as high as the ICsoof an applicable compound or isolated enantiomer disclosed herein.

[0207] The specific dose level and frequency of dosage for any particular subject may be varied and may depend upon a variety of factors, including the activity of the subject compound or isolated enantiomer, the metabolic stability and length of action of that compound or isolated enantiomer, the age, body weight, general health, sex and diet of the subject, mode and time of administration, rate of excretion, drug combination, and severity of the condition of the host undergoing therapy.

[0208] In some embodiments, multiple doses of a compound or isolated enantiomer are administered. The frequency of administration of a compound can vary depending on any of a variety of factors, e.g., severity of the symptoms, condition of the subject, etc. For example, in some embodiments, a compound or isolated enantiomer is administered once per month, twice per month, three times per month, every other week, once per week (qwk), twice per week, three times per week, four times per week, five times per week, six times per week, every other day, daily (qd / od), twice a day (bds / bid), or three times a day (tds / tid), etc.EMBODIMENTS

[0209] Embodiments of the present disclosure include, but are not limited to, the embodiments described in the following clauses:1. An isolated enantiomer of Formula (I):Attorney Reference: GERN-202WO(I) or a pharmaceutically acceptable salt thereof, wherein: n is 0 or an integer from 1 to 5;R1and R2are independently selected from H, alkyl, cycloalkyl, heterocyclyl, and substituted versions thereof, wherein one of R1and R2is absent; and each R3is independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof, or provided that there are two adjacent R3groups, the two adjacent R3groups along with the atoms to which they are attached can form a cyclic ring.2. The isolated enantiomer of clause 1, wherein the isolated enantiomer has the Formula (II):(ID or a pharmaceutically acceptable salt thereof, wherein: n is 0 or an integer from 1 to 4; andA is selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof.3. The isolated enantiomer of clause 2, wherein n is 1 or 2.4. The isolated enantiomer of clauses 2 or 3, wherein R1and R2are independently selected from H and methyl, wherein one of R1and R2is absent.5. The isolated enantiomer of any one of clauses 2-4, wherein each R3is halo.6. The isolated enantiomer of any one of clauses 2-5, wherein:Attorney Reference: GERN-202WO each R4, R5and R6is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof; each R7is independently H or alkyl; and each R8and R9is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof.7. The isolated enantiomer of clause 6, wherein8. The isolated enantiomer of clause 7, wherein each R4is H.9. The isolated enantiomer of clauses 7 or 8, wherein R5and R6are independently selected from H, alkyl and hydroxyl.10. The isolated enantiomer of clause 6, wherein A is.11. The isolated enantiomer of clause 10, wherein R7is H.12. The isolated enantiomer of clauses 10 or 11, wherein each R8is H.13. The isolated enantiomer of any one of clauses 10-12, wherein R9is carboxy or cyano.14. The isolated enantiomer of clause 1, wherein the isolated enantiomer has a structure selected from the following:Attorney Reference: GERN-202WOAttorney Reference: GERN-202WO enantiomer of clause 1, having the following structure: enantiomer of clause 1, having the following structure:enantiomer of clause 1, having the following structure:enantiomer of clause 1, having the following structure:enantiomer of clause 1, having the following structure:H enantiomer of clause 1, having the following structure:Attorney Reference: GERN-202WOg structure: g structure: g structure: g structure: g structure: g structure:Attorney Reference: GERN-202WOg structure: g structure: g structure: g structure: g structure:Attorney Reference: GERN-202WOated enantiomer of clause 1, having the following structure:ated enantiomer of clause 1, having the following structure:ated enantiomer of clause 1, having the following structure:ated enantiomer of clause 1, having the following structure:ated enantiomer of clause 1, having the following structure:Attorney Reference: GERN-202WO37. The isolated enantiomer of clause 1, having the following structure: ntiomer of clause 1, having the following structure:39. The isolated enantiomer of clause 1, having the following structure:40. The isolated enantiomer of clause 1, wherein the isolated enantiomer has a structure selected from the following:Attorney Reference: GERN-202WOAttorney Reference: GERN-202WOAttorney Reference: GERN-202WOg structure: g structure: g structure: g structure:Attorney Reference: GERN-202WOg structure: g structure: g structure: g structure: g structure: g structure:Attorney Reference: GERN-202WOg structure: g structure: g structure: g structure: g structure: g structure:Attorney Reference: GERN-202WOg structure: g structure: g structure: g structure: g structure: g structure:Attorney Reference: GERN-202WOg structure: g structure: g structure: g structure: g structure: g structure:Attorney Reference: GERN-202WO enantiomer of clause 1, having the following structure:enantiomer of clause 1, having the following structure:enantiomer of clause 1, having the following structure:enantiomer of clause 1, having the following structure:enantiomer of clause 1, having the following structure:enantiomer of clause 1, having the following structure:Attorney Reference: GERN-202WOg structure: g structure: g structure: g structure: g structure: g structure:Attorney Reference: GERN-202WOlated enantiomer of clause 1, having the following structure:lated enantiomer of clause 1, having the following structure:lated enantiomer of clause 1, having the following structure:lated enantiomer of clause 1, having the following structure:lated enantiomer of clause 1, having the following structure:Attorney Reference: GERN-202WOlated enantiomer of clause 1, having the following structure:lated enantiomer of clause 1, having the following structure:lated enantiomer of clause 1, having the following structure:lated enantiomer of clause 1, having the following structure:lated enantiomer of clause 1, having the following structure:lated enantiomer of clause 1, having the following structure:Attorney Reference: GERN-202WOg structure: g structure: g structure: g structure: g structure: g structure:Attorney Reference: GERN-202WO98. The isolated enantiomer of clause 1, having the following structure:99. A pharmaceutical composition comprising at least one isolated enantiomer of any one of clauses 1-98 or a pharmaceutically effective salt thereof.100. The pharmaceutical composition of clause 99, further comprising at least one second therapeutic agent.101. The pharmaceutical composition of clause 100, wherein the at least one second therapeutic agent is imetelstat or imetelstat sodium.102. A method of treating a patient for a disease or condition, the method comprising: administering the pharmaceutical composition of any one of clauses 99-101 to the patient.103. The method of clause 102, wherein the disease or condition is a cancer.104. The method of clause 103, wherein the cancer is at least one of hematological malignancies and solid tumors.105. The method of clause 103, wherein the cancer is at least one of acute and chronic leukemias, lymphomas, multiple myeloma, myelodysplastic syndromes (MDS), myeloproliferative neoplasms (MPNs), essential thrombocythemia (ET), polycythemia vera (PV), Chronic Myelogenous Leukemia (CML), myelofibrosis (MF), acute myelogenous leukemia (AML), lung cancers, bladder cancers, liver cancers, colon cancers, adrenal cancers, esophageal cancers, stomach cancers, central nervous system (CNS) cancers, skin cancers, ovarian cancers, cervical cancers, endometrial cancers, renal cancers, prostate cancers, and breast cancers.106. A method of treating a patient for a disease or condition, the method comprising: administering a therapeutically effective amount of at least one isolated enantiomer of any one of clauses 1-98 or a pharmaceutically acceptable salt thereof to the patient.107. The method of clause 106, wherein the disease or condition is a cancer.108. The method of clause 107, wherein the cancer is at least one of hematological malignancies and solid tumors.109. The method of clause 107, wherein the cancer is at least one of acute and chronic leukemias, lymphomas, multiple myeloma, myelodysplastic syndromes (MDS), myeloproliferativeAttorney Reference: GERN-202WO neoplasms (MPNs), essential thrombocythemia (ET), polycythemia vera (PV), Chronic Myelogenous Leukemia (CML), myelofibrosis (MF), acute myelogenous leukemia (AML), lung cancers, bladder cancers, liver cancers, colon cancers, adrenal cancers, esophageal cancers, stomach cancers, central nervous system (CNS) cancers, skin cancers, ovarian cancers, cervical cancers, endometrial cancers, renal cancers, prostate cancers and breast cancers.110. A method of treating a patient for a solid tumor comprising: administering a therapeutically effective amount of at least one compound of Formula (I) to the patient:or a pharmaceutically acceptable salt thereof, wherein: n is 0 or an integer from 1 to 5;R1and R2are independently selected from H, alkyl, cycloalkyl, heterocyclyl, and substituted versions thereof, wherein one of R1and R2is absent; and each R3is independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof, or provided that there are two adjacent R3groups, the two adjacent R3groups along with the atoms to which they are attached can form a cyclic ring.111. The method of clause 110, wherein the at least one compound has the Formula (II):(ID wherein: n is 0 or an integer from 1 to 4; andA is selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof.Attorney Reference: GERN-202WO112. The method of clause 111, wherein n is 1 or 2.113. The method of clauses 111 or 112, wherein R1and R2are independently selected from H and methyl, wherein one of R1and R2is absent.114. The method of any one of clauses 111-113, wherein each R3is halo.115. The method of any one of clauses 111-114, wherein:each R4, R5and R6is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof; each R7is independently H or alkyl; and each R8and R9is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof.116. The method of clause 115, wherein117. The method of clause 116, wherein each R4is H.118. The method of clauses 116 or 117, wherein R5and R6are independently selected from H, alkyl and hydroxyl.119. The method of clause 115, wherein A is.120. The method of clause 119, wherein R7is H.121. The method of clause 119 or 120, wherein each R8is H.122. The method of any one of clauses 119-121, wherein R9is carboxy or cyano.123. The method of clause 110, wherein the at least one compound has a structure selected from:Attorney Reference: GERN-202WOAttorney Reference: GERN-202WOpound has the structure: pound has the structure: pound has the structure: pound has the structure:Attorney Reference: GERN-202WO pound has the structure: pound has the structure: pound has the structure: pound has the structure: pound has the structure: pound has the structure:Attorney Reference: GERN-202WO of clause 110, wherein the at least one compound has the structure:of clause 110, wherein the at least one compound has the structure:of clause 110, wherein the at least one compound has the structure:of clause 110, wherein the at least one compound has the structure:of clause 110, wherein the at least one compound has the structure:of clause 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WOethod of clause 110, wherein the at least one compound has the structure:ethod of clause 110, wherein the at least one compound has the structure:ethod of clause 110, wherein the at least one compound has the structure:ethod of clause 110, wherein the at least one compound has the structure:ethod of clause 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO145. The method of clause 110, wherein the at least one compound has the structure:147. The method of clause 110, wherein the at least one compound has the structure:148. The method of clause 110, wherein the at least one compound has the structure:149. The method of clause 110, wherein the at least one compound has a structure selected from:Attorney Reference: GERN-202WOAttorney Reference: GERN-202WOAttorney Reference: GERN-202WOpound has the structure: pound has the structure: pound has the structure: pound has the structure:Attorney Reference: GERN-202WO pound has the structure: pound has the structure: pound has the structure: pound has the structure: pound has the structure: pound has the structure:Attorney Reference: GERN-202WOd of clause 110, wherein the at least one compound has the structure:d of clause 110, wherein the at least one compound has the structure:d of clause 1101, wherein the at least one compound has the structure:H d of clause 110, wherein the at least one compound has the structure:H d of clause 110, wherein the at least one compound has the structure:d of clause 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO pound has the structure: pound has the structure: pound has the structure: pound has the structure: pound has the structure: pound has the structure:Attorney Reference: GERN-202WO pound has the structure: pound has the structure: pound has the structure: pound has the structure: pound has the structure: pound has the structure:Attorney Reference: GERN-202WO of clause 110, wherein the at least one compound has the structure:of clause 110, wherein the at least one compound has the structure:of clause 110, wherein the at least one compound has the structure:of clause 110, wherein the at least one compound has the structure:of clause 110, wherein the at least one compound has the structure:of clause 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO pound has the structure: pound has the structure: pound has the structure: pound has the structure: pound has the structure: pound has the structure:Attorney Reference: GERN-202WO pound has the structure: pound has the structure: pound has the structure: pound has the structure: pound has the structure:Attorney Reference: GERN-202WO ethod of clause 110, wherein the at least one compound has the structure:ethod of clause 110, wherein the at least one compound has the structure:ethod of clause 110, wherein the at least one compound has the structure:ethod of clause 110, wherein the at least one compound has the structure:ethod of clause 110, wherein the at least one compound has the structure:ethod of clause 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO pound has the structure: pound has the structure: pound has the structure: pound has the structure: pound has the structure:Attorney Reference: GERN-202WO206. The method of clause 110, wherein the at least one compound has the structure: ause 110, wherein the at least one compound has the structure:EXAMPLES

[0210] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the subject matter disclosed herein and are not intended to limit the scope of the present disclosure nor are they intended to represent that the experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric. Standard abbreviations may be used, e.g., bp, base pair(s); kb, kilobase(s); pl, picoliter(s); s or sec, second(s); min, minute(s); h or hr, hour(s); aa, amino acid(s); nt, nucleotide(s); and the like.EXAMPLE 1

[0211] Methods of Making Compounds Useful for Inhibiting Telomerase

[0212] The methods described below are non-limiting examples of methods to make compounds of the present disclosure. A skilled artisan will recognize that there are various modifications that can be performed to make analogs or prepare the compounds in other ways.

[0213] Compound 1: 5-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-5-methyl-3- methylenedihydrofuran-2(3H)-oneAttorney Reference: GERN-202WO5-(4-chloro-2-(1-methyl-1 H-pyrazol-4-yl)phenyl)-5-methyl-3- methylenedihydrofuran-2(3H)-one

[0214] Synthetic scheme:

[0215] Step 1: synthesis of l-(-2-(l-methyl-lH-pyrazol-4-yl)phenyl)ethan-l-one:

[0216] Procedure:

[0217] To a stirred solution of l-(2-bromophenyl)ethan-l-one (1 g, 5.02 mmol) and l-(l-methyl- lH-pyrazol-4-yl)boronic acid (760 mg, 6.02 mmoL) in 1,4-dioxane (20 mL) , potassium carbonate (1.7 g, 12.55 mmol) in water (20 mL) was added. The resulting mixture was degassed for 5 min, and tetrakis(triphenylphosphine)-palladium (500 mg, 10% mol) was added. The reaction mixture was degassed again for further 5 min. The resulting reaction mixture was heated at 70°C for 2 h. The reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was diluted with ethyl acetate (50 mL) and washed with water (2x20 mL). The organic layer was dried over Na2SO4, filtered and the solvent was evaporated under reduced pressure to obtain the crude product. It was purified by column chromatography using silica gel (mesh : 60-120 ) and 40-50% ethyl acetate and petroleum ether as eluent to afford l-(-2-(l-methyl-l / - / -pyrazol-4- yl)phenyl)ethan-l-one as an off-white solid (820 mg, 82%).Attorney Reference: GERN-202WO

[0218] Step 2: Synthesis of methyl 4-hydroxy-4-(2-(l-methyl-lH-pyrazol-4-yl)phenyl)-2-

[0219] Procedure:

[0220] To a stirred solution of l-(-2-(l-methyl-lH-pyrazol-4-yl)phenyl)ethan-l-one (0.5 g, 2.50 mmol) and methyl 2-(bromomethyl)acrylate (580 mg, 3.24 mmoL) in THF (10 mL), Zinc (1.07 g, 16.25 mmol) and saturated NH4CI in water (5 mL) was added. The resulting mixture was stirred at RT for 2 h. The reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was filtered and filtrate was extracted with ethyl acetate (20 mL) and washed with water (2x10 mL). The organic layer was dried over Na2SC , filtered and the solvent was evaporated under reduced pressure to obtain crude product. It was purified by column chromatography using silica gel (mesh: 60-120) and 50-60% ethyl acetate and petroleum ether as eluent to afford methyl 4-hydroxy-4-(2-(l-methyl-lH-pyrazol-4-yl)phenyl)-2-methylenepentanoate as a light brown gum (150 mg, 20%).

[0221] Step 3: Synthesis of 5-methyl-5-(2-(l-methyl-l / 7-pyrazol-4-yl)phenyl)-3-

[0222] Procedure:

[0223] To a stirred solution of methyl 4-hydroxy-4-(2-(l-methyl-l / - / -pyrazol-4-yl)phenyl)-2- methylenepentanoate (150 mg, 0.49 mmol) in DCM (10 mL), trifluoroacetic acid (1 mL) was added and the reaction mixture was stirred at RT for 16 h. The reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was concentrated under reducedAttorney Reference: GERN-202WO pressure to obtain crude product. It was purified by column chromatography using silica gel (mesh : 60-120) and 40-50% ethyl acetate and petroleum ether as eluent to afford 5-methyl-5-(2-(l-methyl- l / - / -pyrazol-4-yl)phenyl)-3-methylenedihydrofuran-2(3H)-one as an off-white gum (64 mg, 47%).

[0224] XH-NMR (400 MHz, CDCI3): 7.73-7.72 (d, 1H), 7.42-7.38(m, 1H), 7.369-7.367 (m, 2H),7.32- 7.28 (m, 1H) 7.168-7.165 (d, 1H), 6.18-6.16 (t, 1H), 5.53-5.52 (t, 1H), 3.97 (s, 3H), 3.10-3.04 (m, 1H), 2.76-2.71 (m, 1H), 1.6 (s, 3H). LCMS: 269.25 (M++H), 99.72% purity.

[0225] 5-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-5-methyl-3-methylenedihydrofuran-2(3H)-one

[0226] TH-NMR (400 MHz, CDCI3): 7.64-7.66 (m, 1H); 7.43 (s, 1H); 7.32-7.36 (m, 2H), 7.17 (d, 1H, 3Hz); 6.17-6.19 (m, 1H); 5.53-5.54 (m, 1H); 3.96 (s, 3H); 3.00-3.05 (m, 1H), 2.70-2.75 (m, 1H); 1.68 (s, 3H) LCMS (M+H): 303.4, 99.93%.

[0227] Compound 2: 4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2- methylenebutanenitrileAttorney Reference: GERN-202WO

[0228] Wherein R1and R2: H, D, Alkyl, cycloakyl. Wherein R1and R2together can make a cyclic ring.

[0229] Step-1: Synthesis of 2-(hydroxymethyl)acrylonitrile:

[0230] Procedure:

[0231] To a solution of diethyl (cyanomethyl)phosphonate (5g, 28.248 mmol) in Water (20 mL) were added Aq.HCHO (3.389 g, 112.992 mmol) and K2CO3 (7.016g, 50.846 mmol) at 0 °C. Reaction mixture was stirred at RT for 3h. Reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was diluted with ether (2X200 mL) and washed with water (2X100 mL). The organic layer was dried over NazSC , filtered, evaporated under reduced pressure, and purified by column chromatography. The desired compound was eluted in 20% ethyl acetate and hexane to afford 2-(hydroxymethyl)acrylonitrile. Yield 2.1g (89%).

[0232] Step-2: Synthesis of 2-(bromomethyl)acrylonitrile:

[0233] Procedure:

[0234] To a stirred solution of 2-(hydroxymethyl)acrylonitrile (2.1g, 25.301 mmol) in Diethyl ether (20 mL) was added PBrs (1.19 ml, 12.65 mmol) at 0 °C. Reaction mixture was stirred at 0 °C for 1 h. Reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was diluted with ether (2X100 mL) and washed with water (2X100 mL). The organic layer was dried over NazSC , filtered, evaporated under reduced pressure, and purified by column chromatography. The desired compound was eluted in 20% ethyl acetate and hexane to afford 2- (bromomethyl)acrylonitrile. Yield 2g (54%).

[0235] Step-3: Synthesis of 2-((phenylsulfonyl)methyl)acrylonitrile:

[0236] Procedure:

[0237] To a stirred solution of 2-(bromomethyl)acrylonitrile (2g, 13.698 mmol) in methanol (20 mL) was added sodium phenyl sulfinate (3.36g, 20.547 mmol) at 0 °C. The reaction mixture wasAttorney Reference: GERN-202WO stirred at 0 °C for 2h. The progress of the reaction was monitored by TLC. Upon disappearance of starting material, the reaction mixture was diluted with ethylacetate (2X100 mL) and washed with water (2X50 mL). The organic layer was dried over NazSO^ filtered, evaporated under reduced pressure and purified by column chromatography. The desired compound was eluted in 40% ethyl acetate in hexane to afford 2-((phenylsulfonyl)methyl)acrylonitrile. Yield 0.62 g (21%).

[0238] Step-4: Synthesis of 2-((tributylsta nnyl)methyl)acrylonitrile:

[0239] Procedure:

[0240] To a stirred solution of 2-((phenylsulfonyl)methyl)acrylonitrile (0.62g, 2.991 mmol) in benzene (6 mL) was added AIBN (0.049g, 0.299 mmol) and tributylstannane (1.306g, 4.487 mmol) at RT. The reaction mixture was stirred at 80 °C for 3 h. The reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was evaporated under reduced pressure and purified by column chromatography. The desired compound was eluted in 10% ethyl acetate and hexane to afford 2-((tributylstannyl)methyl)acrylonitrile. Yield 0.40 g (37%).

[0241] Step-5: Synthesis of 4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2- methylenebutanenitrile:

[0242] Procedure:

[0243] To a stirred solution of 4-chloro-2-(l-methyl-lH-pyrazol-4-yl)benzaldehyde (0.1 g, 0.453 mmol) in benzene (2 mL) was added AIBN (0.0074g, 0.045 mmol) and 2- ((tributylstannyl)methyl)acrylonitrile (0.484 g, 1.359 mmol) at RT. Reaction mixture was stirred at 80 oC for 16h. The progress of the reaction was monitored by TLC. Upon disappearance of starting material, the reaction mixture was evaporated under reduced pressure, and purified by column chromatography. Acetonitrile (6 ml) and concentrated HCI (0.5 ml) were added to the reaction mixture at 0 °C. It was stirred for 10 min, then extracted with Et20 (2X50 ml) and washed with saturated NaHCO3 solution. The organic layer was treated with DBU (0.5 ml) for 10 min. Rection mixture was evaporated under reduced pressure and purified by column chromatography. The desired compound was eluted in 35% Ethyl acetate and hexane to afford 4-(4-chloro-2-(l-methyl- lH-pyrazol-4-yl)phenyl)-4-hydroxy-2-methylenebutanenitrile.

[0244] XH NMR (400 MHz, CDCI3) 67.56 (d, J = 8.4 Hz, 1H), 7.51 (d, J = 2.4 Hz, 2H), 7.36 - 7.27 (m, 2H), 5.95 (s, 1H), 5.78 (s, 1H), 5.21 (dt, J = 8.2, 4.2 Hz, 1H), 3.96 (s, 3H), 2.60 - 2.54 (m, 2H), 2.19 (d, J = 3.7 Hz, 1H). MS (m / z): 288.1 [M+H]+.Attorney Reference: GERN-202WO

[0245] Compound 3: 5-(4-chloro-2-(lH-pyrazol-4-yl)phenyl)-4-methyl-3- methylenedihydrofuran-2(3H)-one5-(4-chloro-2-(1 / - / -pyrazol-4-yl)phenyl)-4-methyl-3-methylenedihydrofuran-2(3H)-one

[0246] Step 1: Synthesis of: l-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-lH-pyrazoleAttorney Reference: GERN-202WO

[0247] Procedure:

[0248] To a stirred solution of 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole (15 g, 77.3 mmol) and 3,4-dihydro-2H-pyran (7.1 g, 85 mmol) in toluene (110 mL), trifluoracetic acid (0.44 g, 0.3 mmol) was added. The reaction mixture was heated to 80°C for 4 h. The progress of the reaction was monitored by TLC (40% ethyl acetate in hexane. Upon completion of starting material, the reaction mixture was allowed to come to room temperature. The reaction mixture was diluted with ethyl acetate and water. The organic layer was separated and washed with water. The organic layer was concentrated under reduced pressure to afford crude l-(tetrahydro-2H-pyran-2-yl)-4- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole and it was used as such for next step.

[0249] Step 2: Synthesis of 4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4- yl)benzaldehyde

[0250] Procedure:

[0251] A suspension of 2-bromo-4-chlorobenazaldehyde (15 g, 68 mmol.), l-(tetrahydro-2H- pyran-2-yl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole (28.5 g, 102.5 mmol.), potassium carbonate (29.7 g, 215.3 mmol.) in 1,4-dioxane (120 mL) and water (30 mL) was degassed for 15 min using nitrogen gas. Subsequently, tetrakis(triphenylphosphine)-palladium (3.9 g, 3.4 mmol.) was added. The reaction mixture was heated to 100°C. The progress of the reaction was monitored by TLC (20% ethyl acetate in hexane). Upon disappearance of starting material, the reaction mixture was allowed to come at room temperature and then it was diluted with ethyl acetate (200 mL) and washed with water (2x100 mL). The organic layer was dried over Na?SO4, filtered and solvent was evaporated under reduced pressure. The crude product thus obtained was purified by column chromatography using silica gel (mesh: 60-120) required compound was eluted at 15% ethyl acetate in hexane to afford 4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4- yl)benzaldehyde (16 g, 81%).Attorney Reference: GERN-202WO

[0252] Step 3: Synthesis of methyl 4-(4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4- yl)phenyl)-4-hydroxy-3-methyl-2-methylenebutanoate

[0253] Procedure:

[0254] To a stirred solution of 4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4- yljbenzaldehyde (16 g, 55 mmol) and methyl-3-bromo-2-methylenebiitanoate (20.1 g, 104.5 mmol) in THF (520 mL), Zinc (6.5 g, 100 mmol) and saturated NH4CI in water (410 mL) were added. The resulting mixture was stirred at room temperature for 2. The progress of the reaction was monitored by TLC (40% ethyl acetate hexane). On completion of starting material, reaction mixture was filtered and diluted with ethyl acetate (600 mL) and washed with brine (2* 300 mL). The organic layer was dried with sodium sulphate, filtered and evaporated under reduced pressure. The crude thus obtained was purified by using column chromatography (silica mesh 60-120) to afford methyl 4-(4-chloro-2- (l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)-4-hydroxy-3-methyl-2-methylenebuta noate as white solid ( yield = 6.6 g, 30%).

[0255] Step 4: Synthesis of 5-(4-chloro-2-(lH-pyrazol-4-yl) phenyl)-4-methyl-3- methylenedihydrofuran-2(3H)-one:

[0256] Procedure:

[0257] To a stirred solution of methyl 4-(4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4- yl)phenyl)-4-hydroxy-3-methyl-2-methylenebutanoate (6.6 g, 16.3 mmol) in 1,4-dioxane (66 mL)Attorney Reference: GERN-202WOHCI (13 mL) was added hydrochloric acid (35 ml, 4N in dioxane) at RT and the reaction mixture was stirred at RT for 6 h. The progress of the reaction was monitored by TLC. Upon disappearance of starting material, the reaction mixture was diluted with ethyl acetate (100 mL) and washed with water (100 mL). The organic layer was washed with sodium bicarbonate solution and brine. The solvent was evaporated under reduced pressure to obtain crude product as brown gum. It was purified by column chromatography (silica mesh 100-200) to afford 5-(4-chloro-2-(lH-pyrazol-4-yl) phenyl)-4-methyl-3-methylenedihydrofuran-2(3H)-one as an off-white gum (2 g, 43%).

[0258] XH-NMR (300 MHz, DMSO);8.02-7.73 (s, 1H), 7.73-7.45 (s, 1H), 7.45 (s, 1H), 7.41-7.38 (d, 1H), 7.21-7.18 (d, 1H), 6.16-6.15 (d, 1H), 6.02-5.99 (d, 1 H), 5.76-5.76 (d, 1H), 3.44 - 3.39 (m, 1H), 0.68-0.66 (d, 3H). LCMS (M+H= 289.1, M-l= 287.2)

[0259] Compound 4: 5-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-3- methylenedihydrofuran-2(3H)-one5-(4-chloro-2-(1-methyl-1 / - / -pyrazol-4-yl)phenyl)-4- hydroxy-3-methylenedihydrofuran-2(3 / - / )-one

[0260] Procedure:

[0261] To a stirred solution of 5-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-3- methylenedihydrofuran-2(3H)-one (lg, 3.2 mmol) in DMSO (5 mL), Selenium dioxide (0.36 g, 3.2 mmol) was added and the reaction mixture was stirred at 80 °C for 1 h. Subsequently, additional amounts of Selenium dioxide (0.36g x 2) was added in 2 portions. Heating was continued for an additional 2 h. The reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was concentrated under reduced pressure to obtain crude product. It was purified by column chromatography using silica gel (mesh : 60-120) and 50-60% ethyl acetate and petroleum ether as eluent to afford 5-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)- 4-hydroxy-3-methylenedihydrofuran-2(3H)-one as an off-white solid (120 mg).Attorney Reference: GERN-202WO

[0262] TH-NMR (400 MHz, CDCI3): 7.61 - 7.53 (m, 2H), 7.36 - 7.33 (m, 1H), 7.30 - 7.26 (m, 1H), 7.23 - 7.21 (m, 1H), 6.50 (bs, 1H), 6.02 (bs, 1H), 5.49 (d, 1H), 4.80 (bs, 1H), 3.97 (s, 3H), 2.97 (d, 1H). MS: 305.0; HPLC: 94.72%.

[0263] Compound 5: 5-(5-chloro-2-(lH-pyrazol-4-yl)phenyl)-3-methylenedihydrofuran-2(3H)- one5-(5-chloro-2-(1 / 7-pyrazol-4-yl)phenyl)-3- methylenedihydrofuran-2(3 / - / )-one

[0264] Synthetic scheme:

[0265] Step-1: Synthesis of l-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-l,3,2-Attorney Reference: GERN-202WO

[0266] Procedure:

[0267] To a stirred solution of 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole (400 mg, 2.68 mmoL) and DHP (248.37 mg, 2.95 mmoL) in toluene (5 mL) , TFA (0.01 mL, 0.13mmoL) was added. The resulting reaction mixture was heated at 80oC for 2 h. The reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was diluted with ethyl acetate (15 mL) and washed with water (2x10 mL). The organic layer was dried over Na2SO4, filtered and then the solvent was evaporated under reduced pressure to obtained l-(tetrahydro- 2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole as a pale yellow oil (600 mg, 75%).

[0268] Step-2: Synthesis of 4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-

[0269] Procedure:

[0270] To a stirred solution of 2-bromo-4-chlorobenzaldehyde (0.3 g, 1.37 mmol) and 1- (tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazolein (226 mg, 1.78 mmoL) in 1,4-dioxane (15 mL), potassium carbonate (500 mg, 3.57 mmol) in water (5 mL) was added. The resulting mixture was degassed for 5 min, and Tetrakis(triphenylphosphine)-palladium (30 mg, 10% mol) was added. The reaction mixture was again degassed for further 5 min. The resulting reaction mixture was heated at 70oC for 2 h. The reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was diluted with ethyl acetate (15 mL) and washed with water (2x10 mL). The organic layer was dried over Na2SO4, filtered and then the solvent was evaporated under reduced pressure. The crude product thus obtained was purified by column chromatography using silica gel (mesh : 60-120) using 25-30% ethyl acetate andAttorney Reference: GERN-202WO petroleum ether as eluent to afford 4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4- yl)benzaldehyde as a pale yellow solid (380 mg, 95%).

[0271] Step-3: Synthesis of methyl 4-(4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-

[0272] Procedure:

[0273] To a stirred solution of 4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4- yl)benzaldehyde (300mg, 1.03 mmol) and methyl 2-(bromomethyl)acrylate (241 mg, 1.34mmoL) in THF (20 mL), Zinc (243 mg, 3.71 mmol) and saturated NH4CI in water (1 mL) was added. The resulting mixture was stirred at RT for 2 h. The reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was filtered and filtrate was extracted with ethyl acetate (20 mL) and washed with water (2x10 mL). The organic layer was dried over Na2SO4, filtered and the solvent was evaporated under reduced pressure to afford methyl-4-(4-chloro-2-(l- (tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2-methylenebutanoate as a colorless gum (350 mg, 86%).

[0274] Step-4: Synthesis of 5-(4-chloro-2-(lH-pyrazol-4-yl)phenyl)-3-methylenedihydrofuran- 2(3H)-one:Attorney Reference: GERN-202WO

[0275] Procedure:

[0276] To a stirred solution of methyl 4-(4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4- yl)phenyl)-4-hydroxy-2-methylenebutanoate (300 g, 0.77 mmol) in DCM (15 mL), trifluro acetic acid (1 mL) was added at RT and the reaction mixture was stirred at RT for 16 h. Reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was concentrated under reduced pressure to obtain brown gum. The crude product was purified by prep-HPLC to afford 5-(4-chloro-2-(lH-pyrazol-4-yl)phenyl)-3-methylenedihydrofuran-2(3H)-oneas an off white gum (15 mg, 6%).

[0277] 1H-NMR (400 MHz, CDCI3): 7.69 (s, 2H), 7.40-7.33 (m, 2H), 6.31-6.30 (t, 1H), 5.68-5.63 (m, 2H), 3.23-3.16 (m, 1H), 2.92-2.84 (m, 1H).

[0278] LCMS (M+H= 275.0, 96.42%).

[0279] 5-(5-chloro-2-(lH-pyrazol-4-yl)phenyl)-3-methylenedihydrofuran-2(3H)-one:

[0280] XH-NMR (400 MHz, CDCI3): 7.67 (s, 2H), 7.445-7.440 (m, 1H), 7.36-7.33 (m, 1H), 7.27-7.24 (m, 1H), 6.33-6.31(t, 1H), 5.69-5.65(m, 1H), 5.64-5.62 (m,lH), , 3.19-3.17 (m, 1H), 2.91-2.89 (m, 1H). LCMS (M+H=275.1, 90.58%)

[0281] Compound 6: 5-(5-fluoro-2-(lH-pyrazol-4-yl)phenyl)-3-methylenedihydrofuran-2(3H)- one5-(5-fluoro-2-(1 / - / -pyrazol-4-yl)phenyl)-3- methylenedihydrofuran-2(3H)-oneAttorney Reference: GERN-202WO

[0282] Synthesis: See synthetic procedure for Compound 5.

[0283] XH-NMR (400 MHz, CDCI3): 7.67 (s, 2H), 7.30-7.27 (m, 1H), 7.18-7.15 (m, 1H), 7.09-7.04 (m, 1H), 6.32-6.30 (t, 1H), 5.68-5.62 (m, 2H), 3.22-3.15 (m, 1H), 2.89-2.61 (m, 1H). LCMS (M+H-259.1, 90.21%)

[0284] Compound 7: 4-(4,5-difluoro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2- methylenebutanenitrile

[0285] Synthesis: See synthetic procedure for Compound 2.

[0286] TH NMR (400 MHz, CDCI3) 6 7.48 - 7.50 (m, 3H), 7.12 - 7.07 (m, 1H), 5.96 (s, 1H), 5.80 (s,1H), 5.21 - 5.15 (m, 1H), 3.96 (s, 3H), 2.60 - 2.54 (m, 2H), 2.15 (d, J = 3.6 Hz, 1H). LCMS (M++H):289.9, 97.62%

[0287] Compound 8: 4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2- methylenebutanoic acidAttorney Reference: GERN-202WO

[0289] Step 1:

[0290] To a stirred solution of 2-bromo-4-chlorobenzaldehyde (400 mg, 1.82 mmoL) and 1- methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole (496 mg, 2.36 mmoL) in 1,4- dioxane (8 mL), potassium carbonate (656 mg, 4.74 mmol) in water (5 mL) was added. The resulting mixture was degassed for 5 min, and tetra kis(triphenylphosphine)-palladium (40 mg, 10 mol%) was added. The reaction mixture was degassed further for 5 min. The resulting reaction mixture was heated at 70°C for 2 h. The reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was diluted with ethyl acetate (15 mL) and washed with water (2x10 mL). The organic layer was dried over NazSC , filtered and then solvent was evaporated under reduced pressure to obtain crude product. It was purified by column chromatography using silica gel (mesh : 60-120 ) and the required compound was eluted at 25-30% ethyl acetate and petroleum ether to afford4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)- lH-pyrazol-4-yl)benzaldehyde as a pale yellow solid (300 mg, 74%).

[0291] Step 2:

[0292] To a stirred solution of 4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4- yl)benzaldehyde (300mg, 1.36 mmol) and methyl 2-(bromomethyl)acrylate (315 mg, 1.77 mmoL) in THF (10 mL), Zinc (320 mg, 4.88 mmol) and saturated NH4CI in water (1 mL) was added. The resulting mixture was stirred at RT for 2 h. The reaction progress was monitored by TLC. Upon disappearanceAttorney Reference: GERN-202WO of starting material, the reaction mixture was filtered and filtrate was extracted with ethyl acetate (20 mL) and washed with water (2x10 mL). The organic layer was dried over NazSCh, filtered and then solvent was evaporated under reduced pressure to afford product methyl 4-(4-chloro-2-(l- methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2-methylenebutanoate as a white gum (280 mg, 65%).

[0293] Step 3:

[0294] To a stirred solution of 4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2- methylenebutanoate (220 mg, 0.68 mmol) in THF water, LiOH. H2O (29 mg, 0.68 mmol) was added at room temperature. Reaction mixture was stirred for 2 h. Reaction mixture was concentrated and residue was taken in water (10 mL) . Aqueous layer was washed with Ethyl acetate (2 x 10 mL). Aqueous layer was acidified with citric acid and extracted with Ethyl acetate (2 x 10 mL). Combined organic extract were dried over Na?SO4, filtered and concentrated to afford target compound (180 mg)

[0295] 1H-NMR (400 MHz, CDCI3): 12.4-12.2 (brs, 1H), 7.93 (s, 1H), 7.58 (s, 2H), 7.30-7.29 (m, 2H), 6.076-6.072 (d, 1H), 5.59 (s, 1H), 5.3-5.2 (brs, 1H), 3.99 (s, 3H), 2.56-2.51 (m, 1H), 2.28 (m, 1H).LCMS: m / z 307.1 [M++H]; (92.99% purity)

[0296] Compound 9: 5-(4-chloro-2-(lH-pyrazol-4-yl)phenyl)-3-methylenedihydrofuran-2(3H)- one5-(4-chloro-2-(1 H-pyrazol-4-yl)phenyl)-3- methylenedihydrofuran-2(3 / - / )-oneAttorney Reference: GERN-202WO

[0297] Synthetic scheme:

[0298] Step 1: Synthesis of l-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-l,3,2-

[0299] Procedure:

[0300] To a stirred solution of 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole (400 mg, 2.68 mmoL) and DHP (248.37 mg, 2.95 mmoL) in toluene (5 mL) , TFA (0.01 mL, 0.13mmoL) was added. The resulting reaction mixture was heated at 8O0C for 2 h. The reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was diluted with ethyl acetate (15 mL) and washed with water (2x10 mL). The organic layer was dried over Na2SO4, filtered and then the solvent was evaporated under reduced pressure to obtained l-(tetrahydro- 2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole as a pale yellow oil (600 mg, 75%).

[0301] Step 2: Synthesis of 4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4- yl)benzaldehyde:Attorney Reference: GERN-202WO

[0302] Procedure:

[0303] To a stirred solution of 2-bromo-4-chlorobenzaldehyde (0.3 g, 1.37 mmol) and 1- (tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazolein (226 mg, 1.78 mmoL) in 1,4-dioxane (15 mL), potassium carbonate (500 mg, 3.57 mmol) in water (5 mL) was added. The resulting mixture was degassed for 5 min, and Tetrakis(triphenylphosphine)-palladium (30 mg, 10% mol) was added. The reaction mixture was again degassed for further 5 min. The resulting reaction mixture was heated at 70oC for 2 h. The reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was diluted with ethyl acetate (15 mL) and washed with water (2x10 mL). The organic layer was dried over Na2SO4, filtered and then the solvent was evaporated under reduced pressure. The crude product thus obtained was purified by column chromatography using silica gel (mesh : 60-120) using 25-30% ethyl acetate and petroleum ether as eluent to afford 4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4- yl)benzaldehyde as a pale yellow solid (380 mg, 95%).

[0304] Step 3: Synthesis of methyl 4-(4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-Attorney Reference: GERN-202WO

[0305] Procedure:

[0306] To a stirred solution of 4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4- yljbenzaldehyde (300mg, 1.03 mmol) and methyl 2-(bromomethyl)acrylate (241 mg, 1.34mmoL) in THF (20 mL), Zinc (243 mg, 3.71 mmol) and saturated NH4CI in water (1 mL) was added. The resulting mixture was stirred at RT for 2 h. The reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was filtered and filtrate was extracted with ethyl acetate (20 mL) and washed with water (2x10 mL). The organic layer was dried over Na2SO4, filtered and the solvent was evaporated under reduced pressure to afford methyl-4-(4-chloro-2-(l- (tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2-methylenebutanoate as a colorless gum (350 mg, 86%).

[0307] Step 4: Synthesis of 5-(4-chloro-2-(lH-pyrazol-4-yl)phenyl)-3-methylenedihydrofuran- 2(3H)-one:

[0308] Procedure:

[0309] To a stirred solution of methyl 4-(4-chloro-2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-4- yl)phenyl)-4-hydroxy-2-methylenebutanoate (300 g, 0.77 mmol) in DCM (15 mL), trifluro acetic acid (1 mL) was added at RT and the reaction mixture was stirred at RT for 16 h. Reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was concentrated under reduced pressure to obtain brown gum. The crude product was purified by prep-HPLC to afford 5-(4-chloro-2-(lH-pyrazol-4-yl)phenyl)-3-methylenedihydrofuran-2(3H)-oneas an off-white gum (15 mg, 6%).

[0310] 1H-NMR (400 MHz, CDCI3): 7.69 (s, 2H), 7.40-7.33 (m, 2H), 6.31-6.30 (t, 1H), 5.68-5.63 (m, 2H), 3.23-3.16 (m, 1H), 2.92-2.84 (m, 1H). LCMS (M+H= 275.0, 96.42%).Attorney Reference: GERN-202WO

[0311] Compound 10a (R)-4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2- methylenebutanoic acid and Compound 10b (S)-4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)- 4-hydroxy-2-methylenebutanoic acid:Compound 10a Compound 10b

[0312] Synthetic protocol: (R)-4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2- methylenebutanoic acid and (S)-4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2- methylenebutanoic acid

[0313] Step 1: Synthesis of 5-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-3- methylenedihydrofuran-2(3H)-one:

[0314] To a stirred solution of 4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2- methylenebutanoate (1 g, 3.1 mmol) in DCM (15 mL), trifluroacetic acid (1 mL) was added at RT and the reaction mixture was stirred at RT for 16 h. Reaction progress was monitored by TLC. Upon disappearance of starting material, the reaction mixture was concentrated under reduced pressure to obtain brown gum. The crude product was purified by silica gel column using 5% Methanol DCM to afford 5-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-3- methylenedihydrofuran-2(3H)-one as an off white solid (870 mg). The compound was further purified using preparative chiralpak IG HPLC column to isolate both isomers: lsomer-1: 380 mg and lsomer-2: 410 mg.Attorney Reference: GERN-202WO

[0315] Step 2 (Synthesis of Compound 10a): To a stirred solution of 4-(4-chloro-2-(l-methyl-lH- pyrazol-4-yl)phenyl)-3-methylenedihydrofuran-2(3H)-one (isomer-1), (380 mg, 0.17 mmol) in THF water, LiOH. H2O (29 mg, 0.68 mmol) was added at room temperature. Reaction mixture was stirred for 2 h. Reaction mixture was concentrated and residue was taken in water (10 mL) . Aqueous layer was washed with Ethyl acetate (2 x 10 mL). Aqueous layer was acidified with citric acid and extracted with Ethyl acetate (2 x 10 mL). Combined organic extract were dried over NazSC)4, filtered and concentrated to afford target compound (180 mg)

[0316] XH-NMR (400 MHz, CDCI3): 12.4-12.2 (brs, 1H), 7.93 (s, 1H), 7.58 (s, 2H), 7.30-7.29 (m, 2H), 6.076-6.072 (d, 1H), 5.59 (s, 1H), 5.3-5.2 (brs, 1H), 3.99 (s, 3H), 2.56-2.51 (m, 1H), 2.28 (m, 1H). LCMS: m / z 307.1 [M++H]; HPLC purity : 99.03% , Chiral purity: 99.78% (Chiral pack IG, 20% Ethanol: 0.2% DEA in Hexane)

[0317] Step 3 (Synthesis of Compound 10b): To a stirred solution of 4-(4-chloro-2-(l-methyl-lH- pyrazol-4-yl)phenyl)-3-methylenedihydrofuran-2(3H)-one (isomer-2), (410 mg, 0.14 mmol) in THE water, LiOH. H2O (29 mg, 0.68 mmol) was added at room temperature. Reaction mixture was stirred for 2 h. Reaction mixture was concentrated and residue was taken in water (10 mL) . Aqueous layer was washed with Ethyl acetate (2 x 10 mL). Aqueous layer was acidified with citric acid and extracted with Ethyl acetate (2 x 10 mL). Combined organic extract were dried over Na2SO4, filtered and concentrated to afford target compound (265 mg)

[0318] 1H-NMR (400 MHz, CDCI3): 12.4-12.2 (brs, 1H), 7.93 (s, 1H), 7.58 (s, 2H), 7.30-7.29 (m, 2H), 6.076-6.072 (d, 1H), 5.59 (s, 1H), 5.3-5.2 (brs, 1H), 3.99 (s, 3H), 2.56-2.51 (m, 1H), 2.28 (m, 1H). LCMS: m / z 307.1 [M++H]; HPLC purity : 98.73% , Chiral purity: 99.97% (Chiral pack IG, 20% Ethanol: 0.2% DEA in Hexane)

[0319] Note: Stereochemistry in both enantiomers has been tentatively assigned.

[0320] Compound 11: 5-(4,5-dichloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-methyl-3- methylenedihydrofuran-2(3H)-oneAttorney Reference: GERN-202WO

[0321] Synthetic schemeK CO Pd(Ph3P)4

[0323] Procedure:

[0324] To a suspension of 2-bromo-4,5-dichlorobenazaldehyde (10 g, 39 mmol.), l-methyl-4- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole (12.2 g, 59 mmol.) and potassium carbonate (17.1 g, 124 mmol.) in 1,4-dioxane (100 mL) was degassed for 15 min using nitrogen gas. To the reaction mixture, tetrakis (triphenylphosphine)-palladium (2.27 g, 1.9 mmol.) was added. The reaction mixture was heated to 100°C. The progress of the reaction was monitored by TLC (70% ethyl acetate in hexane). Upon disappearance of the starting material, the reaction mixture was cooled to room temperature and then diluted with ethyl acetate (200 mL) and washed with water (2x100 mL). The organic layer was dried over NazSCh. It was filtered and then solvent was evaporated under reduced pressure to obtain crude residue. It was purified by column chromatography using silica gel (mesh: 60-120) and eluted with 40% ethyl acetate in hexane to obtain 4,5-dichloro-2-(l-methyl-lH-pyrazol- 4-yl)benzaldehyde (5 g, 50% ).

[0325] Step-2: Synthesis of methyl 4-(4,5-dichloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4- hydroxy-3-methyl-2-methylenebuta noate:Attorney Reference: GERN-202WO

[0326] Procedure:

[0327] To a stirred solution of 4,5-dichloro-2-(l-methyl-lH-pyrazol-4-yl)benzaldehyde (2.5 g, 9.8 mmol) and methyl-3-bromo-2-methylenebutanoate (3.4 g, 17mmoL) in THF (80 ml_). Zinc (1.6 g, 24 mmol) and saturated NH4CI in water (60 mL) were added. The resulting mixture was stirred at RT for 2 h. The reaction progress was monitored by TLC (70% ethyl acetate hexane). Upon disappearance of starting material, the reaction mixture was filtered and the filtrate was extracted with ethyl acetate (150 mL) and washed with water (2x100 mL). The organic layer was dried over NazSCU, filtered and solvent was evaporated under reduced pressure to afford methyl-4-(4,5- dichloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-3-methyl-2-methylenebuta noate as a colorless gum (3 g, 83%).

[0328] Step 3: Synthesis of 5-(4,5-dichloro-2-(lH-pyrazol-4-yl) phenyl)-4-methyl-3- methylenedihydrofuran-2(3H)-one:

[0329] Procedure:

[0330] To a stirred solution of methyl 4-(4,5-dichloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4- hydroxy-3-methyl-2-methylenebutanoate (2 g, 0.77 mmol) in DCM (20 mL), trifluoro acetic acid (6 mL) was added at RT. The reaction mixture was stirred at RT for 8 h. The progress of the reaction was monitored by TLC using ethyl acetate as eluent. Upon disappearance of starting material, the reaction mixture was concentrated under reduced pressure to obtain the crude product. It wasAttorney Reference: GERN-202WO purified by column chromatography (silica mesh 100-200) to afford 5-(4,5-dichloro-2-(l-methyl-H- pyrazol-4-yl) phenyl)-4-methyl-3-methylenedihydrofuran-2(3H)-one (0.6 g, 33%).

[0331] 1H-NMR (300 MHz, DMSO):8.02 (s, 1H), 7.71 (s, 1H), 7.66 (s, 1H), 7.38 (s, 1H), 6.18-6.18 (d, 1H), 6.00-5.97 (d, 1H), 5.80 (d, 1H), 3.47 - 3.42 (m, 1H), 3.89 (s, 3H), 0.70-0.67(d, 3H). LCMS (M+H+): 337.5, 97.08%.

[0332] Compound 12: 4-(4-chloro-2-(l-(2-hydroxyethyl)-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2- methylenebutanenitrile

[0334] Procedure:

[0335] Step-1: Synthesis of 4-chloro-2-(l-(2-hydroxyethyl)-lH-pyrazol-4-yl)benzaldehyde

[0336] 2-bromo-4-chlorobenzaldehyde (2.40 g, 10.96 mmol) was dissolved in a mixture of 1,4- dioxane and water (4:1, v / v) under stirring at room temperature. To this solution were added 2-(4- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazol-l-yl)ethan-l-ol (3.91 g, 16.40 mmol), potassium carbonate (K2CO3) (4.54 g, 32.8 mmol), and [Pd(PPh3)4] (0.13 g, 0.11 mmol) under nitrogen atmosphere. The reaction mixture was stirred at 100 °C for 8 hours. The progress of the reaction was monitored by TLC using ethyl acetate: hexane (8:2) as the mobile phase. Upon completion, the reaction mixture was cooled to room temperature, poured into cold water, and extracted with ethyl acetate. The combined organic layers were dried, filtered, and concentrated under reduced pressure. The crude product thus obtained was purified by columnAttorney Reference: GERN-202WO chromatography, by using 55% ethylacetate and hexane as eluent to afford 4-chloro-2-(l-(2- hydroxyethyl)-lH-pyrazol-4-yl)benzaldehyde as a white solid in 83% yield.

[0337] MS (ESI): m / z = 251.2 [M+H]+

[0338] Step-2: Synthesis of 4-(4-chloro-2-(l-(2-hydroxyethyl)-lH-pyrazol-4-yl)phenyl)-4-hydroxy- 2-methylenebutanenitrile

[0339] 4-chloro-2-(l-(2-hydroxyethyl)-lH-pyrazol-4-yl)benzaldehyde (2.80 g, 11.19 mmol) was dissolved in a THF:water mixture (1:1, v / v) and stirred at room temperature. To this solution were added 2-(bromomethyl)acrylonitrile (4.90 g, 33.59 mmol) and indium powder (1.542 g, 13.40 mmol). The reaction mixture was stirred overnight at room temperature under ambient atmosphere. Completion of the reaction was confirmed by TLC using ethyl acetate: hexane (8:2) as the eluent. The mixture was then filtered through a celite pad, diluted with water, and extracted with ethyl acetate. The combined organic extracts were dried, filtered, and concentrated under reduced pressure. The crude product thus obtained was purified by column chromatography by using 55% ethylacetate and hexane as eluent to afford 4-(4-chloro-2-(l-(2-hydroxyethyl)-lH- pyrazol-4-yl)phenyl)-4-hydroxy-2-methylenebutanenitrile as a solid in 60% yield.

[0340] 1H NMR (300 MHz, CDCI3): 6 7.58-7.55 (m, 3H), 7.36-7.28 (m, 2H), 5.91 (s, 1H), 5.77 (s, 1H), 5.29-5.22 (m, 1H), 4.31-4.28 (m, 2H), 4.03 (s, 2H), 3.09 (s, 1H), 2.55-2.51 (m, 2H), 2.19-2.18 (m, 1H).

[0341] MS (ESI): m / z = 317 [M+H]+

[0342] Compound 13: 4-(4-chloro-2-(l-(tetrahydro-2H-pyran-4-yl)-lH-pyrazol-4-yl)phenyl)-4- hydroxy-2-methylenebutanenitrile4-(4-chloro-2-(1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)phenyl)-4-hydroxy-2- methylenebutanenitrileAttorney Reference: GERN-202WO

[0343] Synthetic scheme

[0344] Procedure:

[0345] Step-1: Synthesis of 4-chloro-2-(l-(tetrahydro-2H-pyran-4-yl)-lH-pyrazol-4- yl)benzaldehyde

[0346] A solution of 2-bromo-4-chlorobenzaldehyde (1.30 g, 5.93 mmol) in 1,4-dioxane and water (4:1, v / v) was stirred at room temperature under a nitrogen atmosphere. To this solution were added l-(tetrahydro-2H-pyran-4-yl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH- pyrazole (2.47 g, 8.90 mmol), potassium carbonate (K2CO3) (2.45 g, 17.80 mmol), and [Pd(PPh3)4] (0.14 g, 0.12 mmol). The reaction mixture was then heated to 100 °C and stirred for 8 hours. Reaction progress was monitored by TLC using ethyl acetate:hexane (7:3) as the eluent. Upon completion, the reaction mixture was cooled to ambient temperature, poured into cold water, and extracted with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The crude residue was purified by column chromatography by using 45% ethylacetate and hexane as eluent. The desired product 4-chloro-2-(l-(tetrahydro-2H-pyran-4-yl)-lH-pyrazol-4-yl)benzaldehyde (3) was obtained as a solid in 87% yield.

[0347] MS (ESI): m / z = 291.2 [M+H]+

[0348] Step-2: Synthesis of 4-(4-chloro-2-(l-(tetrahydro-2H-pyran-4-yl)-lH-pyrazol-4-yl)phenyl)- 4-hydroxy-2-methylenebutanenitrile

[0349] 4-chloro-2-(l-(tetrahydro-2H-pyran-4-yl)-lH-pyrazol-4-yl)benzaldehyde (3.90 g, 13.40 mmol) was dissolved in a THE : water mixture (1:1, v / v) by stirring at room temperature. To the resulting solution, 2-(bromomethyl)acrylonitrile (4.89 g, 33.50 mmol) and indium powder (1.85 g, 16.09 mmol) were added. The reaction mixture was stirred overnight at room temperature under ambient conditions. Reaction progress was monitored by TLC using ethyl acetate:hexane (8:2) as the eluent. Upon completion, the reaction mixture was filtered through a Celite pad, and the filtrate was diluted with water. The aqueous layer was extracted with ethyl acetate, and theAttorney Reference: GERN-202WO combined organic layers were separated. The aqueous layer was concentrated under reduced pressure to dryness. The residue was purified by column chromatography by using 35% ethylacetate and hexane as eluent, to afford the final compound as a white solid in 69% yield.

[0350] 1H NMR (300 MHz, CDCI3): 6 7.61-7.53 (m, 3H), 7.35-7.28 (m, 2H), 5.93 (s, 1H), 5.77 (s, 1H), 5.22-5.18 (m, 1H), 4.36 (s, 1H), 4.14-4.08 (m, 2H), 3.69-3.49 (m, 2H), 2.64-2.49 (m, 1H), 2.11-2.03 (m, 6H).

[0351] MS (ESI): m / z - 358.2 [M+H]+

[0352] Compound 14: 4-(4-chloro-2-(l-(2-oxo-2-(piperidin-l-yl)ethyl)-lH-pyrazol-4-yl)phenyl)-4- hydroxy-2-methylenebutanenitrile4-(4-chloro-2-(1 -(2-oxo-2-(piperidin-1 - yl)ethyl)-1 / 7-pyrazol-4-yl)phenyl)-4- hydroxy-2-methylenebutanenitrile

[0353] Synthetic scheme

[0354] Procedure:

[0355] Step-1: Synthesis of tert-butyl 2-(4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH- pyrazol-l-yl)acetateAttorney Reference: GERN-202WO

[0356] 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole (10.00 g, 51.0 mmol) was dissolved in DMF and stirred at room temperature. To this solution were added tert-butyl 2- bromoacetate (13.99 g, 71.70 mmol) and potassium tert-butoxide (KOtBu) (6.44 g, 57.0 mmol). The reaction mixture was stirred at room temperature for 2 hours, and reaction progress was monitored by TLC using 55% ethylacetate and hexane as eluent. Upon completion, the reaction was quenched with water and extracted with ethyl acetate. After purification using column chromatography by using 15% ethylacetate and hexane as eluent, the desired product tert-butyl 2- (4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazol-l-yl)acetate was obtained as gummy solid in 89% yield.

[0357] 1H NMR (300 MHz, CDCI3): 6 7.82 (s, 1H), 7.76 (s, 1H), 4.83 (s, 2H), 1.47 (s, 9H), 1.31 (s, 12H).

[0358] Step-2: Synthesis of tert-butyl 2-(4-(5-chloro-2-formylphenyl)-lH-pyrazol-l-yl)acetate

[0359] 2-bromo-4-chlorobenzaldehyde (3.5 g, 15.0 mmol) was dissolved in a mixture of 1,4- dioxane and water (4:1, v / v) and stirred at room temperature under a nitrogen atmosphere. To the reaction mixture were added tert-butyl 2-(4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH- pyrazol-l-yl)acetate (7.38 g, 23.0 mmol), potassium carbonate (K2CO3) (6.61 g, 47.8 mmol), and [Pd(PPh3)4] (250 mg, 0.217 mmol). The reaction mixture was heated to 100 °C and stirred for 8 hours. Reaction progress was monitored by thin-layer chromatography (TLC) using 10% ethyl acetate in hexane as the eluent. Upon completion, the mixture was cooled to room temperature. Water (50 ml) was added to the reaction mixture and it was extracted with ethyl acetate. After concentration of the organic layer and purification by column chromatography by using 4% methanol and DCM as eluent, the product was obtained as a white solid in 64% yield.

[0360] 1H NMR (300 MHz, CDCI3): 6 10.17 (s, 1H), 7.92 (d, J = 8.4 Hz, 1H), 7.69 - 7.63 (m, 2H), 7.46 (d, J = 1.8 Hz, 1H), 7.42 - 7.33 (m, 1H), 4.89 (s, 2H), 1.50 (s, 9H).

[0361] Step-3: Synthesis of 2-(4-(5-chloro-2-formylphenyl)-lH-pyrazol-l-yl)acetic acid

[0362] tert-butyl 2-(4-(5-chloro-2-formylphenyl)-lH-pyrazol-l-yl)acetate (2.20 g, 6.80 mmol) was dissolved in 20 mL of DCM and stirred at room temperature. To this solution, TFA (2 mL) was added dropwise, and the reaction mixture was stirred at room temperature for 2 hours. Reaction progress was monitored by TLC using 30% ethyl acetate in hexane as the eluent. Upon completion, reaction mixture was evaporated to dryness and was used without further purification in the next step.Attorney Reference: GERN-202WO

[0363] 1H NMR (300 MHz, DMSO-d6): 6 10.10 (s, 1H), 8.17 (s, 1H), 7.88 - 7.85 (m, 2H), 7.65 (s, 1H), 7.55 - 7.51 (m, 1H), 5.03 (s, 2H).

[0364] Step-4: Synthesis of 4-chloro-2-(l-(2-oxo-2-(piperidin-l-yl)ethyl)-lH-pyrazol-4- yljbenzaldehyde

[0365] To a stirred solution of 2-(4-(5-chloro-2-formylphenyl)-lH-pyrazol-l-yl)acetic acid 13 (6.00 g, 23.0 mmol) in DMF, EDC (6.51 g, 33.95 mmol), HOBt (4.60 g, 33.95 mmol), and DIPEA (11.87 mL, 68.18 mmol) were added sequentially under an inert atmosphere. After complete dissolution, piperidine (4.07 g, 47.0 mmol) was added to the reaction mixture. The reaction mixture was stirred overnight at room temperature, and the reaction progress was monitored by thin-layer chromatography (TLC) using pure ethyl acetate as the eluent. The reaction mixture was poured in cold water, extracted with ethyl acetate and purified with column chromatography by using 80% ethylacetate and hexane as eluent to obtain desired compound in 75% yield.

[0366] 1H NMR (300 MHz, DMSO-d6): 6 10.18 (s, 1H), 7.91 (d, J = 8.4, 1H), 7.71 - 7.60 (m, 2H), 7.51 (s, 1H), 7.46 - 7.41 (m, 1H), 5.06 (s, 2H), 3.61-3.42 (m, 5H).

[0367] Step-5: Synthesis of 4-(4-chloro-2-(l-(2-oxo-2-(piperidin-l-yl)ethyl)-lH-pyrazol-4- yl)phenyl)-4-hydroxy-2-methylenebutanenitrile

[0368] 4-chloro-2-(l-(2-oxo-2-(piperidin-l-yl)ethyl)-lH-pyrazol-4-yl)benzaldehyde (1.70 g, 5.10 mmol) was dissolved in a THF:water mixture (1:1, v / v) and stirred at room temperature. To the reaction mixture, were added 2-(bromomethyl)acrylonitrile (2.04 g, 12.80 mmol) and indium powder (0.71 g, 6.1 mmol). The reaction mixture was stirred overnight at room temperature, and the reaction progress was monitored by TLC using pure ethyl acetate as the eluent. Upon completion, the mixture was filtered through a Celite pad, followed by the addition of water. The aqueous phase was extracted with ethyl acetate, and the organic and aqueous layers were separated. The organic layer was evaporated under reduced pressure to dryness. The crude product was purified by column chromatography by using 2% methanol in dichloromethane as eluent to afford the final compound as gummy solid in 58% yield.

[0369] JH NMR ( CDCI3400 MHz): 7.65 (s, 1H), 7.55 - 7.53 (m, 2H), 7.32 - 7.31 (m, 2H), 5.92 (s, 1H), 5.79 (s, 1H), 5.20 - 5.17 (m, 1H), 5.02 (s, 2H), 3.55 (t, J = 5.6Hz, 2H), 3.47 - 3.45 (m, 2H), 2.57 - 2.55 (m, 2H), 2.00 (s, 1H), 1.67 - 1.66 (m, 2H), 1.61 - 1.56 (m,4H).

[0370] MS (ESI): m / z = 398.8 [M]+Attorney Reference: GERN-202WO

[0371] Compound 15: 4-(4-chloro-2-(l-(2-morpholino-2-oxoethyl)-lH-pyrazol-4-yl)phenyl)-4- hydroxy-2-methylenebutanenitrile4-(4-chloro-2-(1 -(2-morpholino-2- oxoethyl)-1 / - / -pyrazol-4-yl)phenyl)-4- hydroxy-2-methylenebutanenitrile

[0372] Synthetic scheme

[0373] Procedure:

[0374] Step-1: Synthesis of 4-chloro-2-(l-(2-morpholino-2-oxoethyl)-lH-pyrazol-4- yl)benzaldehyde

[0375] To a stirred solution of 2-(4-(5-chloro-2-formylphenyl)-lH-pyrazol-l-yl)acetic acid (1.50 g, 5.68 mmol) in DMF, EDC (1.63 g, 8.52 mmol), HOBt (1.30 g, 8.52 mmol), and DIPEA (2.20 g, 17.00 mmol) were added sequentially at room temperature under inert atmosphere. After 15 minutes of activation, morpholine (0.49 g, 5.68 mmol) was added to the reaction mixture. The mixture was stirred overnight at room temperature, and the reaction progress was monitored by TLC using ethyl acetate as the eluent. The reaction mixture was poured in cold water, extracted with ethyl acetate and purified with column chromatography by using 80% ethylacetate and hexane as eluent to obtain desired compound in 72% yield.

[0376] MS (ESI): m / z = 333.9 [M+H]+

[0377] Step-2: Synthesis of 4-(4-chloro-2-(l-(2-morpholino-2-oxoethyl)-lH-pyrazol-4-yl)phenyl)- 4-hydroxy-2-methylenebutanenitrile

[0378] 4-chloro-2-(l-(2-morpholino-2-oxoethyl)-lH-pyrazol-4-yl)benzaldehyde (1.435 g, 4.30 mmol) was dissolved in a THF:water mixture (1:1, v / v) and stirred at room temperature. To thisAttorney Reference: GERN-202WO solution were added 2-(bromomethyl)acrylonitrile (0.941 g, 6.45 mmol) and indium powder (0.592 g, 5.16 mmol). The reaction mixture was stirred overnight at room temperature, and the reaction progress was monitored by TLC using 5% methanol in dichloromethane as the eluent. After completion, the mixture was filtered through a celite pad, followed by the addition of water. The aqueous phase was extracted with ethyl acetate, and the organic and aqueous layers were separated. The aqueous layer was evaporated to dryness under reduced pressure. The crude residue was purified by column chromatography by using 2% methanol in dichloromethane as eluent, to afford the final compound as a yellowish solid in 77% yield.

[0379] 1H NMR (400 MHz, CDCI3): 67.65 (s, 1H), 7.55-7.54 (m, 2H), 7.33-7.30 (m, 2H), 5.91 (s, 1H), 5.78 (s, 2H), 5.18-5.14 (m, 1H), 5.00 (s, 2H), 3.70-3.66 (m, 4H), 3.60-3.46 (m, 4H), 2.56-2.53 (m, 2H).

[0380] MS (ESI): m / z = 400.7 [M]+

[0381] Compound 16: 4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-3,4-dihydroxy-2- methylenebutanenitrile4-(4-chloro-2-(1 -methyl-1 H-pyrazol-4- yl)phenyl)-3,4-dihydroxy-2- methylenebutanenitrile

[0382] Synthetic scheme

[0383] Procedure:

[0384] Step-1: Synthesis of 4-chloro-2-(l-methyl-lH-pyrazol-4-yl)benzaldehydeAttorney Reference: GERN-202WO

[0385] 2-bromo-4-chlorobenzaldehyde (2.50 g, 11.4 mmol) was dissolved in a dioxane:water mixture (4:1, v / v) and stirred at room temperature under a nitrogen atmosphere. To this solution were added l-methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole (3.5 g, 17.1 mmol), potassium carbonate (K2CO3) (4.7 g, 34.2 mmol), and [Pd(PPh3)4] (131.8 mg, 0.114 mmol). The reaction mixture was heated to 100 °C and stirred for 8 hours. Reaction progress was monitored by thin-layer chromatography (TLC) using ethyl acetate: hexane (7:3) as the eluent. After completion, the reaction mixture was cooled to room temperature, poured into cold water, and extracted with ethyl acetate. The combined organic layers were dried and concentrated under reduced pressure. The crude product was purified by column chromatography by using 25% ethylacetate and hexane as eluent to afford the target compound 4-chloro-2-(l-methyl-lH-pyrazol- 4-yl)benzaldehyde as a white solid in 67% yield.

[0386] (ESI): m / z = 220.9 [M+H]+

[0387] Step-2: Synthesis of 4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2- methylenebutanenitrile

[0388] 4-chloro-2-(l-methyl-lH-pyrazol-4-yl)benzaldehyde (0.200 g, 0.0009 mole) was dissolved in THF:H2O (1:1) mixture by stirring at room temperature. 2-(bromomethyl)acrylonitrile (0.1326 g, 0.0009 mole) and indium (0.1043 g, 0.0009 mole) were added to the reaction mixture and continue the stirring overnight till the completion of the reaction on TLC (7:3, ethylacetate : hexane as eluent). The reaction mixture was filtered through celite band followed by addition of water to the reaction mixture. The aqueous layer was extracted with ethyl acetate. The layers were separated, and the aqueous layer was evaporated to dryness. The target compound 4-(4-chloro-2- (l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2-methylenebutanenitrile was obtained in 78% yield after purification with column chromatography by using 35% ethylacetate and hexane as eluent.

[0389] MS (ESI): m / z = 287.8 [M+H]+

[0390] Step-3: Synthesis of 4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-3,4-dihydroxy-2- methylenebutanenitrile

[0391] 4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2-methylenebutanenitrile (0.100 mg, 0.347 mole) was dissolved in DMSO with stirring. SeCh (0.57 mg, 0.5206 mole) was added in portions and temperature was raised to 100 C for 5 h. The stirring was continued till the completion of the reaction on TLC (7:3, ethylacetate : hexane as eluent). The reaction mixture was filtered through celite band followed by addition of water to the reaction mixture. The aqueousAttorney Reference: GERN-202WO layer was extracted with ethyl acetate. The layers were separated, and the aqueous layer was evaporated to dryness. The target compound 4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-3,4- dihydroxy-2-methylenebutanenitrile was obtained in 12% yield after purification with column chromatography by using 35% ethylacetate and hexane as eluent.

[0392] 1H NMR (400 MHz, CDCI3): 6 7.70 (s, 1H), 7.61 - 7.59 (m, 1H), 7. 54 - 7.52 (m, 1H), 7.37 - 7. 31 (m, 2H), 6.07 (s, 1H), 5.96 (s, 1H), 5.16 (d, J = 7.2, 1H), 4.33 (d, J = 7.2, 1H), 3.94 (s, 3H).

[0393] MS (ESI): m / z = 304.09 [M + H]+

[0394] Compound 17: 4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-2-methylene-4- oxobutanenitrile4-(4-chloro-2-(1-methyl-1 / - / -pyrazol-4- yl)phenyl)-2-methylene-4-oxobutanenitrile

[0395] Synthetic scheme

[0396] Procedure:

[0397] Step-1: Synthesis of 4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-2-methylene-4- oxobutanenitrile

[0398] 4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2-methylenebutanenitrile (1.0 g, 3.47 mmol) was dissolved in dichloromethane (DCM) and stirred at room temperature. Dess- Martin periodinane (DMP) (2.94 g, 6.95 mmol) was added portion-wise to the reaction mixture over several minutes. The reaction was monitored by TLC using 5% methanol in DCM as the eluent, and was found to be complete after 4 hours. Upon completion, 50 mL of DCM was added, and the reaction mixture was washed sequentially with aqueous sodium thiosulfate and then with water. The organic layer was separated, dried, and concentrated under reduced pressure to afford theAttorney Reference: GERN-202WO crude product. The purification was done using column chromatography by using 40% ethylacetate and hexane as eluent to afford 4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-2-methylene-4- oxobutanenitrile in 76% yield.

[0399] 1H NMR (400 MHz, CDCI3): 67.60-7.59 (m, 1H), 7.47 (s, 1H), 7.42-7.41 (m, 1H), 7.37-7.31 (m, 2H), 6.90-6.04 (m, 1H), 5.69-5.68 (m, 1H), 3.72-3.62 (m, 3H), 3.18 (s, 2H).

[0400] MS (ESI): m / z = 285.07 [M]+

[0401] Compound 18 (2S,4S)-4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2- methylbutanenitrile and Compound 19 (2 / ?,4S)-4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4- hydroxy-2-methylbutanenitrile:(2S,4S)-4-(4-chloro-2-( 1 -methyl- (2R,4S)-4-(4-chloro-2-(1-methyl-1 / - / -pyrazol-4-yl)phenyl)-4- 1 / - / -pyrazol-4-yl)phenyl)-4-hydroxy- hydroxy-2-methylbutanenitrile 2-methylbutanenitrile

[0402] Synthetic scheme

[0403] Procedure:

[0404] Step-1: Synthesis of 4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2- methylbutanenitrile

[0405] (S)-4-(4-chloro-2-(l-methyl-lH-pyrazol-4-yl)phenyl)-4-hydroxy-2-methylenebutanenitrile (200 mg, 0.69 mmol), obtained via chiral column chromatography, was dissolved in methanol (MeOH) and stirred at room temperature. To this solution, palladium on carbon (Pd / C, 10%) (40 mg) was added, and a hydrogen balloon was attached to the reaction vessel. The reaction mixture was stirred under a hydrogen atmosphere for 1 hour, and the reaction progress was monitored by TLC using ethyl acetate:hexane (7:3) as the eluent. Upon completion, the mixture was filtered through a Celite pad, and the filtrate was concentrated under reduced pressure. The crudeAttorney Reference: GERN-202WO products were purified by column chromatography by using 25% ethylacetate and hexane as eluent to afford the desired hydrogenated compounds and the diasteromers were separated using column chromatography (stereochemistry after hydrogenation was tentatively assigned).

[0406] Compound 18:XH NMR (CDCI3400 MHz): 7.90 (s, 1H), 7.60 - 7.53 (m, 2H), 7.37 - 7.35 (m, 1H), 7.30 - 7.25 (m, 2H), 5.55 (d, J = 4Hz, 1H), 4.93 - 4.90 (m, 1H), 3.86 (s, 3H), 3.04 - 2.99 (m, 1H), 1.98 - 1.67 (m, 2H), 1.24 - 1.15 (m, 2H). MS (ESI): m / z = 290.0 [M+H]+

[0407] Compound 19:XH NMR (CDCI3400 MHz): 7.91 (s, 1H), 7.59 - 7.53 (m, 2H), 7.38 - 7.35 (m, 1H), 7.28 - 7.25 (m, 2H), 5.45 (d, J = 4Hz, 1H), 4.85 - 4.80 (m, 1H), 3.89 (s, 3H), 2.91 - 2.82 (m, 1H), 1.98 - 1.67 (m, 1H), 1.24 - 1.15 (m, 3H). MS (ESI): m / z = 290.0 [M+H]+EXAMPLE 2

[0408] Isolation of Enantiomers

[0409] Racemates of each of Compounds 1, 2, 3, 4, 12, 13, 14, and 15 were subjected to chiral separation methods to isolate each enantiomer.

[0410] The enantiomeric and / or isomeric forms of the compounds were resolved using high- performance liquid chromatography (HPLC) as described in Methods 1-7 below. Separations were carried out on chiral stationary phase columns, with column dimensions and specific conditions provided individually for each compound. The mobile phases typically comprise n-hexane:ethanol, with either 0.1% diethylamine (DEA) or trifluoroacetic acid (TFA) added as a modifier, depending on the nature of the compound, to enhance peak resolution and minimize tailing. Parameters including flow rate, detection wavelength, and column temperature are detailed in the respective methods.

[0411] Each method resulted in baseline separation of all isomers, with retention times (RTs) recorded for each isomer. These RTs were consistent across multiple runs, demonstrating the robustness and reproducibility of the chromatographic procedures.

[0412] The structural assignments of the isomers were made based on theoretical considerations, and are tentatively represented in the accompanying figures. The absolute stereochemical configurations of the isomers of Compound 2 and Compound 4 were confirmed via X-ray crystallography.

[0413] Chiral HPLC separation methods:

[0414] General description of methods:Attorney Reference: GERN-202WO

[0415] Sample preparation

[0416] For each of Methods 1-7 described herein, about 50 mg of the subject compound was weighed in a 5 ml dried conical flask. About 5 ml of EtOH was added to the flask. The contents were then sonicated for 1 - 2 minutes to make sure that the compound dissolved completely, and the sample solution was clear. The sample was filtered through a 0.45 pm syringe filter.

[0417] Method 1:

[0418] In Method 1, enantiomers of the subject compound were separated using samples prepared as described above on which chiral HPLC was performed in accordance with the following parameters:

[0419] Method 2:

[0420] In Method 2, enantiomers of the subject compound were separated using samples prepared as described above on which chiral HPLC was performed in accordance with the following parameters:Attorney Reference: GERN-202WO

[0421] Method 3:

[0422] In Method 3, enantiomers of the subject compound were separated using samples prepared as described above on which chiral HPLC was performed in accordance with the following parameters:

[0423] Method 4:

[0424] In Method 4, enantiomers of the subject compound were separated using samples prepared as described above on which chiral HPLC was performed in accordance with the following parameters:Attorney Reference: GERN-202WO

[0425] Method 5:

[0426] In Method 5, enantiomers of the subject compound were separated using samples prepared as described above on which chiral HPLC was performed in accordance with the following parameters:

[0427] Method 6:

[0428] In Method 6, enantiomers of the subject compound were separated using samples prepared as described above on which chiral HPLC was performed in accordance with the following parameters:Attorney Reference: GERN-202WO

[0429] Method 7:

[0430] In Method 7, enantiomers of the subject compound were separated using samples prepared as described above on which chiral HPLC was performed in accordance with the following parameters:

[0431] Separation of isomers of Compound 3:

[0432] This compound was separated using Method 6.

[0433] Peak-1 (Compound 3a): RT 7.85min; Chiral HPLC purity: 99.12%

[0434] Peak-2 (Compound 3b): RT 9.14min; Chiral HPLC purity: 99.61%

[0435] Separation of isomers of Compound 1:

[0436] This compound was separated using Method 5.

[0437] Peak-1 (Compound la): RT 5.27min; Chiral HPLC purity: 100%

[0438] Peak-2 (Compound lb): RT 10.14min; Chiral HPLC purity: 99.89%

[0439] Separation of isomers of Compound 4:

[0440] This compound was separated using Method 4.

[0441] Peak-1 (Compound 4a): RT 4.70min; Chiral HPLC purity: 99.92%

[0442] Peak-2 (Compound 4b): RT 8.35min; Chiral HPLC purity: 99.95%

[0443] Separation of isomers of Compound 12:

[0444] This compound was separated using Method 1.

[0445] Peak-1 (Compound 12a): RT 16.35min; Chiral HPLC purity: 99.88%Attorney Reference: GERN-202WO

[0446] Peak-2 (Compound 12b): RT 21.25min; Chiral HPLC purity: 99.49%

[0447] Separation of isomers of Compound 13:

[0448] This compound was separated using Method 2.

[0449] Peak-1 (Compound 13a): RT 8.94min; Chiral HPLC purity: 99.43%

[0450] Peak-2 (Compound 13b): RT 11.34min; Chiral HPLC purity: 99.01%

[0451] Separation of isomers of Compound 15:

[0452] This compound was separated using Method 3.

[0453] Peak-1 (Compound 15a): RT 35.18min; Chiral HPLC purity: 98.86%

[0454] Peak-2 (Compound 15b): RT 44.25min; Chiral HPLC purity: 93.31%

[0455] Separation of isomers of Compound 14:

[0456] This compound was separated using Method 3.

[0457] Peak-1 (Compound 14a): RT 12.54min; Chiral HPLC purity: 99.26%

[0458] Peak-2 (Compound 14b): RT 16.37min; Chiral HPLC purity: 95.32%

[0459] Separation of isomers of Compound 2:

[0460] This compound was separated using Method 7.

[0461] Peak-1 (Compound 2a): RT 9.25min; Chiral HPLC purity: 99.62%

[0462] Peak-2 (Compound 2b): RT 11.79min; Chiral HPLC purity: 99.33%EXAMPLE 3

[0463] Proliferation Assay

[0464] 10000 cells / well were seeded in a 96-well assay plate. Four different types of cells were seeded separately containing both telomerase positive cell lines viz., MOLM-13 and HL-60, and telomerase negative cells lines viz., Saos-2 and MRC-9, respectively (see Table 1 and Table 2). The required concentrations of the compounds, the reference compounds, and the positive control (doxorubicin) were added at 6-point concentrations starting from 100 pM, followed by 1 / 3 dilutions in duplicates. After 72 hours of incubation, cell-titer gio reagent was added directly to each well.Attorney Reference: GERN-202WOLuminescence read was measured 10 minutes after incubation. Doxorubicin (10 pM) was used as a positive control for cell death. Untreated cells were used as controls to calculate IC50 values based on no inhibition / cell death observed in absence of treatment.

[0465] Annexin V: Apoptosis Detection Assay

[0466] 0.2xl0A6 cells / well were plated in a 96-well plate. Compounds were added at 6-point dose-response curves starting from lOOuM, followed by 1 / 10 dilutions in duplicates. After 72 hours incubation, RealTime-Glo™Annexin V Apoptosis reagent was added and luciferase activity was measured as a readout for apoptosis.

[0467] Telomere Length Screening Assay

[0468] Relative Human Telomere Length is quantified using the Quantification qPCR Assay Kit (cat No 8908, ScienCell). The average telomere length of the samples is directly compared across samples. The telomere primer set recognizes and amplifies the telomere sequence. For normalization, a single copy reference (SCR) primer set that recognizes and amplifies a 100 bp-long region on human chromosome 17 is used as a reference. The average length of chromosomal telomeres in a sample is confirmed by the ratio (T / S) of the copy number of telomere gene (Tel) and the copy number of single copy gene (SCR).

[0469] Telomerase Repeated Amplification Protocol (TRAP) Assay

[0470] The telomeric repeat amplification protocol consists of three main stages: primer elongation, amplification of telomerase-synthesized DNA, and thirdly, its detection, which is analogous to telomerase activity.

[0471] Telomerase Enzyme Assay

[0472] 0.2xl0A6 cells / well were seeded in a 6-well plate. Heat-inactivated cells were used as a negative control. The cells were incubated for 48 hours and then collected to determine telomerase activity using the TeloTAGGG Telomerase PCR ELISAPLUS kit (Roche, UK) (see Table 1). Untreated cells were used as a positive control. Untreated cells treated with RNAse and heat inactivated were used as a negative control.Attorney Reference: GERN-202WO

[0473] Mouse Pharmacokinetic (PK) Assay

[0474] The pharmacokinetic (PK) profiles of test compounds were obtained at 10 mg / kg by oral (PO) route in mice (BALB / c, male mice). Results from the mouse PK studies are summarized in Table 2.

[0475] See data for IC50, selectivity and mouse pharmacokinetics (PK) in Table 1 and Table 2 below.Table 1. IC50 valuesAttorney Reference: GERN-202WOTable 2. Selectivity and Mice PKAttorney Reference: GERN-202WOAttorney Reference: GERN-202WO

[0476] Pharmacokinetic (PK) profiles in mice

[0477] PK profiles for Compounds 1-4 at 50 mg / kg per dose by oral (PO) route are shown in FIGS. 1-4, respectively.

[0478] The PK profile of Compound 1 at 10 mg / kg per dose by oral (PO) route in NOD-SCID mice is shown in Table 5 and FIG. 5. Acceptable plasma exposure levels were demonstrated at the tested dose by PO route.Table 5. PK profile of Compound 1.Attorney Reference: GERN-202WOAs used herein, "BLQ" means "below the limit of quantification."

[0479] The PK profile of Compound 3 at 10 mg / kg by oral (PO) route in NOD-SCID mice is shown in Table 6 and FIG. 6. Acceptable plasma exposure levels were demonstrated at the tested dose by PO route. Concentrations were observed in tissues up to 24 hours (FIG. 7).Table 6. PK profile of Compound 3.

[0480] The PK profile of Compound 5 at 10 mg / kg by oral (PO) route in NOD-SCID mice is shown in Table 6 and FIG. 8. Acceptable plasma exposure levels were demonstrated at the tested dose byPO route.Table 7. PK profile of Compound 5.Attorney Reference: GERN-202WOEXAMPLE 4

[0481] In vivo pilot efficacy study using MOLM13-LUC cell line and 10 mpk doses

[0482] Study design is shown in FIG. 9. All groups (control and compound treated; see Table 3) were inoculated with IM MOLM13-Luc cells via i.v. in a tail vein (one animal was kept as blank for imaging). The animals were randomized on Day 8 and dosed orally at 10 mpk every day starting on Day 8 through Day 28. Bioluminescence imaging for tumor was evaluated on Days 0, 10, 15, 18, 21, 25 and 28. Body weight was monitored every day before dosing. BIBR-1532 was dosed at 25mpk daily. The following compounds were tested in this efficacy study: Group 1 (Compound 3); Group 2 (Compound 10b); Group 3 (Compound 11); Group 4 (Compound 2); Group 5 (BIBR-1532); and Group 6 (vehicle control).

[0483] Bioluminescence imaging (BLI): The optimized concentration of cells and signal was monitored by Luminescence imaging in vitro followed by intravenous injections to the animals with the required concentrations. Animals were peritoneally injected with 150mg / kg D-luciferin potassium salt before undergoing anaesthesia with 3% isoflurane in air in an anaesthesia induction box. Then 2% isoflurane in air / 02 was continuously delivered via a nose cone system in the dark box of a high sensitivity CCD camera using the I VIS® Spectrum In vivo Imaging System from PerkinElmer. BLI was performed 5 minutes after substrate injection. Mice were placed in the dorsal, lateral and ventral positions followed by Acquisition. Sequential (2-3) images were taken for a set of 5 animals, one animal per group (as only 5 animals can be accommodated in the instrument), after 5 mins of D-Luciferin injection.Attorney Reference: GERN-202WO

[0484] Bioluminescence imaging for tumor evaluation was done on Days 0, 5, 10, 15, 18 and 21 respectively along with regular body weight monitoring.

[0485] Compounds tested:Compound 2 Compound 3 Compound 10b Compound 11BIBR-1532Table 3. Study animal groups.

[0486] Results and Discussion.

[0487] Based on this in vivo efficacy study on MOLM13-Luc cells and using 10 mpk doses, the following conclusions were drawn. Group 1 and Group 4 showed a maximum regression of tumour load as observed via bioluminescence imaging levels. See FIGS. 19-22. Group 4 showed no mortality as of Day 40 (FIG. 23). Consistent body weights were seen across groups throughout the study (FIG. 24). The animals in Group 4 showed good overall health with no mortality. Group 1 and Group 2 showed extended survival time compared to BIBR-1532. BIBR-1532 showed 50% mortality through Day-40.Attorney Reference: GERN-202WOEXAMPLE 5

[0488] In vivo efficacy study using MOLM13-LUC cell line and 50 mpk doses

[0489] Study design is shown in FIG. 10. MOLM-13-Luc cells were cultured and collected at the log phase and suspended in DPBS. Each mouse (NOD / SCID mice) was intravenously injected in a tail vein with tumor cells in a total volume of 0.1 mL in DPBS at 1 million concentration (based on the in vivo pilot study for MOLM13-Luc cells set forth in Example 4). The inoculation day was denoted as Day 0. The cell viability was checked before and after cell inoculation. The study lasted for 21-28 days or extended further based on overall health of the animals. The study included 5 groups (7 animals per group); wherein each group was dosed with one of the following: Vehicle, one of 3 compounds, or BIRB-1532 as a reference compound. The test compounds were dosed at 50 mg / kg, and BIRB-1532 also at 50 mg / kg, on Day 10 after inoculation of cells.

[0490] The following compounds were tested in this efficacy study: Group 3 (vehicle control); Group 4 (Compound 9); Group 5 (Compound 4); Group 6 (Compound 1); and Group 7 (Bl BR-1532).

[0491] The peaks of BLI were read and images of tumor progression monitored from Day 10. This was in reference to the pilot study in selecting the concentration of cells and the day of dosing based on tumor burden. The animals were dosed every alternate day until the end of the study (subject to change based on the overall health of the animals). The clinical evaluation upon termination included analysis of all images (from Day 0, Day 5, Day 10, Day 15, Day 17, Day 21 and Day 27), the percent survival vs the vehicle, the body weight of the animals, and evaluations of markers across tissue specimens. The tissues collected from the treated and the vehicle group included brain, lungs, eyes, lymph nodes and blood. The tissues were collected and frozen for any analysis in future based on the study outcome. A parallel satellite PK analysis with 3 animals per test compound was also performed.

[0492] All groups (see Table 4) were inoculated with IM MOLM13-Luc cells, except for Groups 1 and 2, which were kept as blank and single compound treatment, respectively. The dosing regimen was modified as per the animal health condition and has been described below:

[0493] Day-8: 50 mpk (Groups 4, 5, 6 and 7).

[0494] Day-10 and Day-12: 25 mpk for Groups 4 and 6, 10 mpk for Group 5, and 50 mpk for Group 7.

[0495] Day-17 onwards (alternate day): 10 mpk (Groups - 4, 5 and 6) and 50 mpk for Group 7.Attorney Reference: GERN-202WO

[0496] Bioluminescence imaging: The optimized concentration of cells and signal was monitored by Luminescence imaging in vitro followed by intravenous injections to the animals with the required concentrations. Animals were peritoneally injected with 150mg / kg D-luciferin potassium salt before undergoing anaesthesia with 3% isoflurane in air in an anaesthesia induction box. Then 2% isoflurane in air / 02 was continuously delivered via a nose cone system in the dark box of a high sensitivity CCD camera using the I VIS® Spectrum In vivo Imaging System from PerkinElmer. BLI was performed 5 minutes after substrate injection. Mice were placed in the dorsal, lateral and ventral positions followed by Acquisition. Sequential (2-3) images were taken for set of 5 animals, one animal per group (as only 5 animals can be accommodated in the instrument) after 5 mins of D- Luciferin injection.

[0497] Bioluminescence imaging for tumor evaluation was done on Days 0, 5, 10, 15, 18 and 21, respectively, along with regular body weight monitoring.

[0498] Compounds tested:Compound 1 Compound 4 Compound 9BIBR-1532Table 4. Study animal groups.Attorney Reference: GERN-202WO

[0499] Results and Discussion.

[0500] Based on this in vivo efficacy study on MOLM13-Luc cells and using 50 mpk doses, Group 6 showed a maximum regression of tumour load as observed via bioluminescence imaging levels and overall health of the animals from this group. Group 6 showed maximum regression and better profile than BIBR-1532. Based on the 28-day luminescence signal evaluation, Group 5 was comparable to BIBR-1532 at a lower dose. See FIGS. 11-12. For Groups 4, 5 and 6, weight loss was observed initially, followed by stabilization of body weight (FIG. 13). A few animals in Groups 4, 5 and 6 were lost early in the study due to overdosing (FIG. 14). Tumor regression is most prominently observed in Group 6 (FIGS. 15-17).EXAMPLE 6

[0501] 4-day acute toxicity study in BALB / c mice

[0502] A 4-day Gavage Maximum Tolerability Study (MTD) was performed in BALB / c mice using various compounds of the present disclosure. The study design is shown in FIG. 18. A summary of the results is described below.

[0503] Compound 1

[0504] The oral formulation of Compound 1 was tolerable up to 50 mg / kg for 4 days in BALB / c mice. No abnormal clinical signs were observed at doses 10 mg / kg, 25mg / kg and 50 mg / kg. Normal body weight was observed in all three test groups. No mortality was observed in any of the test dose groups. Normal whole blood values were observed in all treatment groups.

[0505] Histopathology of Compound 1

[0506] Mild to moderate inflammatory infiltrations were observed in lungs and kidney tissues at 10, 25, and 50 mg / kg. However, the brain appears to have been unaffected by the treatments, as its histopathology remained normal in all groups, including the control (vehicle-treated) group.Attorney Reference: GERN-202WO

[0507] Compound 4

[0508] The oral formulation of Compound 4 was tolerable up to 50 mg / kg for 4 days in BALB / c mice. No abnormal clinical signs were observed at doses 10 mg / kg, 25mg / kg and 50 mg / kg. All test groups exhibited normal increments in body weight, comparable to the group receiving the vehicle control. No mortality was observed from any of the test dose groups. Normal whole blood values were observed in all treatment groups.

[0509] Histopathology of Compound 4

[0510] Mild to moderate inflammatory infiltrations were noted in the tissues of the heart, lungs, and kidneys at doses of 10, 25, and 50 mg / kg. Additionally, mild congestion and infiltration were observed in the blood vessels of the heart and kidneys. The spleen exhibited a reduction in the pulp area, while the brain seemed unaffected by the treatment. Mild congestion of the central vein was observed in the liver.

[0511] Compound 2a

[0512] The oral administration of Compound 2a was well tolerated up to 50 mg / Kg over a 4-day period in BALB / c mice. No abnormal clinical signs were observed at the administered test doses of 10 mg / kg, 25 mg / kg, and 50 mg / kg. All test groups exhibited normal increments in body weight, comparable to the group receiving the vehicle control. No mortality was observed in any of the vehicle and test dosed groups. Normal whole blood values were observed in all treatment groups.

[0513] Histopathology of Compound 2a

[0514] Based on histopathological observation, no toxic reactions (degeneration, necrosis, inflammatory and hyperplasic changes) were observed in spleen, heart, brain, in three test groups (G2, G3 and G4) when compared with control group animals. As shown in FIG. 18, G2 animals received doses of Compound 2a at 10 mg / kg; G3 animals received doses of Compound 2a at 25 mg / kg; and G4 animals received doses of Compound 2a at 50 mg / kg. Foci of infiltration of inflammatory cells were observed in centri lobular / peri portal region of the liver in two out of three [2 / 3] animals from the G1 and G2 groups and one out of three [1 / 3] animals from the G4 group. But the G3 group did not show such changes in liver. Alveolar / interstitial wall thickening or inflammation was observed in the lung in two out of three [2 / 3] animals from the G1 group and one out of three [1 / 3] animals from the G2 and G4 groups. But the G3 group did not show such changes in the lung. Tubular / interstitial infiltration / inflammation was observed in one out of three [1 / 3] animals from the G3 group, but the other groups Gl, G2 and G4 did not show such changes inAttorney Reference: GERN-202WO kidneys. No toxic or treatment related changes were observed in the G2, G3 and G4 groups. Very few changes in liver and lung were observed in the G1 control group, which may be attributed to spontaneous changes.

[0515] Results for Compound 2a

[0516] The study results indicated that the oral administration of Compound 2a to BALB / c mice at doses of 10 mg / kg, 25 mg / kg, and 50 mg / kg demonstrated good tolerability. The absence of observable adverse effects, such as body weight reduction and mortality, at these dosage levels indicated a favorable safety profile for Compound 2a in the tested mice. There were no notable abnormalities in any of the blood parameters, indicating that the drug, at the specified doses, does not have a pronounced adverse impact on these specific blood parameters. Histopathological examinations of the brain, heart, kidney, liver, lung, and spleen showed no differences between the vehicle control and the Compound 2a groups at doses of 10, 25, and 50 mg / kg. Therefore, oral administration of Compound 2a at doses up to 50 mg / kg body weight was determined to be nontoxic and deemed safe.EXAMPLE 7

[0517] Further profiling of enantiomers using assays

[0518] Screening and assay details:

[0519] I. Cell viability assay: Protocol

[0520] Assay optimization: 500, 1000, 2000 and 5000 Cells were plated in 96-well plate in duplicate and cultured in complete media for 72 hours. After 72 hours, cell titer-glo reagent was added and luminescence was measured using a plate reader. Luminescence units were plotted against cell-number and an optimal cell number was selected for inhibitor screening based on curve linearity and signal to noise ration.

[0521] Inhibitor screening: At day 0, optimized cell number was plated in a 96-well plate in duplicates. After 24 hours incubation, cells were treated with various concentrations of the inhibitor (100uM-0.411uM, 6-point DRC in duplicates). After 72 hours, an effect on cell proliferation was measured through cell titer-glo reagent. Doxorubicin (@10uM) was used as positive control (Six wells, 100% cells death) and untreated cells as negative controls (six wells). BIBR-1532 was used as the reference compound. Overall, six test compounds and one tool compound (BIBR-1532) wereAttorney Reference: GERN-202WO screened per-96 well plate. Various parameters such as Z', IC50 values, slope and R2values were generated by plotting the raw data in graph-pad software.

[0522] Selectivity assessment: Telomerase negative Saos-2 osteosarcoma cells were used for selectivity against cancer cells and MRC-9 lung fibroblast cells for selectivity against normal human cells. The test compound should ideally show no-inhibition or very high IC50 compared to the test or tool compounds (Final criteria were fixed after analyzing the activity of early set of test compounds).

[0523] II. Telomerase activity assay: Protocol

[0524] To investigate whetherthe inhibitor treatment reduced telomerase activity, the enzymatic activity was measured by Telo TAGGG telomerase PCR ELISA kit (Roche Diagnostics GmbH, Mannheim, Germany) according to the manufacturer's protocol.

[0525] In the first step, telomerase added telomeric repeats (TTAGGG) to the 3' end of the biotin- labeled synthetic primer. In a second step, these elongation products were amplified by PCR, generating PCR products with the telomerase-specific 6 nucleotide increments. As opposed to other TRAP assay formats, the TeloTAGGG Telomerase PCR ELISA contained all compounds required for the telomerase reaction and PCR in a ready-to-use reaction buffer for combining both reactions in one-step / one-tube-reaction. An additional advantage over the conventional assay was the use of optimized primer sequences eliminating the need for hot-start PCR or separation of the primers by a wax barrier, and avoided amplification artifacts, such as primer dimers.

[0526] In brief, at day 0, 2xl05cells were plated in a 6-well cell culture plate and 24 hours later treated with selected concentrations of the inhibitor (10uM-0.041uM, 6-point DRC). After incubation for 72 hours, the cells were washed with cold PBS, lysed in 200 pl lysis buffer and incubated on ice for 30 min. The lysate was centrifuged at 16,000xg at 4 °C for 20 min. Later the supernatant was collected, and the protein extracts were subjected to TRAP assay. For the TRAP reaction, 25 pl of reaction mixture, 10 pg of sample, and nuclease free water to make a final volume of 50 pl. PCR was performed, and ELISA was carried out following the manufacturer's guidelines. A total of 5 pl of PCR products were added to a streptavidin-coated 96-well plate in duplicates and hybridized to a digoxigenin (DIG)-labeled telomeric repeat-specific detection probe. The immobilized PCR products were detected with peroxidase-conjugated anti-DIG antibody. After addition of the stop reagent, the plate was assessed on a plate reader at a wavelength of 450 nmAttorney Reference: GERN-202WO within 30 min. The cell extracts were heat inactivated for 10 min at 85 °C and used as negative controls.

[0527] In each 96-well ELISA plate, 7 compounds including the tool compound can be screened.

[0528] III. Apoptosis assay: Protocol

[0529] Flow cytometry assay was performed by Annexin V / PI double staining method to evaluate the induction of apoptosis. At day 0, 2xl05cells were plated in a 6-well cell culture plate and 24 hours later treated with selected concentrations of the inhibitor. After incubation for 72 hours, the cells were dissociated using accutase and resuspended at ~1 x io6cells / mL in binding buffer and add lOOuL / well to a polypropylene 96-well plate in duplicates from each 6-well plate. 5 pL of FITC Annexin V and 10 pL of Propidium Iodide Solution were added and incubated for 20 minutes at room temperature. 400 pl IX Binding buffer was added to each tube and gently mixed. The cells were analyzed immediately (within 1 hour) by flow cytometry.

[0530] Apoptosis / necrotic cells (Pl+ / Annexin V+) and viable cells (Pl- / Annexin V-) were quantified through flow cytometry based on the staining pattern. Apoptotic cells were detected and expressed as the percentage of total cells.ReagentsAttorney Reference: GERN-202WO

[0531] IV. CYP inhibition assay: Protocol

[0532] Objective: To determine Inhibition potential of test compounds against CYP 3A4 isoform at 10 pM concentration using Human liver microsomes.

[0533] Assay Procedure: The working stock solution (1.0 pL) was added to 179 pL of the isoformspecific cocktail mixture and pre-incubated at 37°C for 10 minutes in duplicates. Following preincubation, 20 pL of pre-warmed NADPH was added, and the mixture was incubated further in a shaking water bath as per the protocol summary. After incubation, the reactions were quenched with 200 pL of an ice-cold solution containing an isotope-labeled internal standard. The samples were then vortexed at 1000 rpm for 10 minutes. Blank and Blank + IS samples were prepared by adding 179 pL of the cocktail mixture and 1 pL of vehicle, followed by 200 pL of ice-cold acetonitrile (for Blank) or 200 pL of internal standard (for Blank + IS). These samples were vortexed for 10 minutes, and 20 pL of NADPH was subsequently added and mixed for 30-60 seconds. All samples were centrifuged at 4000 rpm for 30 minutes. After centrifugation, 100 pL of the supernatant was diluted with 100 pL of water (1:1), vortexed for 5 minutes, and analyzed by LC-MS / MS.Attorney Reference: GERN-202WO

[0534] Results from each of the assays profiling the enantiomers described above are shown inTable 8, Table 9, Table 10, Table 11, Table 12, Table 13, Table 14 and Table 15 below.Table 8: Profiling of Enantiomers of Compound 2Table 9: Profiling of Enantiomers of Compound 1Attorney Reference: GERN-202WOTable 10: Profiling of Enantiomers of Compound 4Attorney Reference: GERN-202WOTable 11: Profiling of Enantiomers of Compound 3Attorney Reference: GERN-202WOTable 12: Profiling of Enantiomers of Compound 12Table 13: Profiling of Enantiomers of Compound 13Attorney Reference: GERN-202WOTable 14: Profiling of Enantiomers of Compound 15Attorney Reference: GERN-202WOTable 15: Profiling of Enantiomers of Compound 14EXAMPLE 8

[0535] Equilibrium solubility (24h) of the crystalline form of Compound 2a at different pH conditions

[0536] Study protocol:

[0537] Caffeine (1 mg / mL in pH 6.8 buffer) and Compound 2a (1 mg / mL in pH 1.2, 4.5, and 6.8 buffers) were weighed considering purity and molecular weight.

[0538] Samples were incubated for 24 hours at room temperature on a thermomixer at 1200 rpm.

[0539] Post incubation, samples were centrifuged at 4000 rpm for 10 minutes. The supernatants were collected and analyzed using LCMS, with a 7-point calibration curve ranging from 3.12 pg / mL to 200 pg / mL.Attorney Reference: GERN-202WO

[0540] Results:Attorney Reference: GERN-202WOEXAMPLE 9 - Crystallization and Characterization of Single Crystals of Isolated Enantiomers Materials and Methods

[0541] Compound la

[0542] Single crystals of Compound la were obtained by slow evaporation of an ethyl acetate solution at room temperature. A suitable crystal was selected under a polarizing microscope and mounted on a XtaLAB Synergy, Dualflex, HyPix diffractometer. Data collection was carried out at 298 K. The structure was solved using Intrinsic Phasing with the SHELXT program [2] and refined using full-matrix least-squares minimization on F2with the SHELXL refinement package [3], all implemented within the Olex2 software suite [1], All non-hydrogen atoms were refined anisotropically. See FIG. 29.

[0543] Compound 4a

[0002] Single crystals of Compound 4a were obtained by slow evaporation of acetonitrile solution of compound at room temperature. A suitable crystal was selected under a polarizing microscope and mounted on a XtaLAB Synergy, Dualflex, HyPix diffractometer. Data collection was carried out at 100 K. Using Olex2 [1], the structure was solved using Intrinsic Phasing with the SHELXT program [2] and refined using full-matrix least-squares minimization on F2with the SHELXL refinement package [3], All non-hydrogen atoms were refined anisotropically. See FIG. 27.

[0544] Compound 4b

[0003] Single crystals of Compound 4b were obtained by slow evaporation of acetonitrile solution of compound at room temperature. A suitable crystal was selected under a polarizing microscope and mounted on a XtaLAB Synergy, Dualflex, HyPix diffractometer. Data collection was carried out at 100 K. Using Olex2 [1], the structure was solved using Intrinsic Phasing with the SHELXT program [2] and refined using full-matrix least-squares minimization on F2with the SHELXL refinement package [3], All non-hydrogen atoms were refined anisotropically. See FIG. 28.

[0545] Compound 2a

[0004] Single crystals of Compound 2a were obtained by slow evaporation of dichloromethane solution of compound at room temperature. A suitable crystal was selected under a polarizing microscope and mounted on a XtaLAB Synergy, Dualflex, HyPix diffractometer. Data collection was carried out at 100 K. Using Olex2 [1], the structure was solved using Intrinsic Phasing with the SHELXTAttorney Reference: GERN-202WO program [2] and refined using full-matrix least-squares minimization on F2with the SHELXL refinement package [3], All non-hydrogen atoms were refined anisotropically. See FIG. 25.

[0546] Compound 2b

[0005] Single crystals of Compound 2b were obtained by slow evaporation of dichloromethane solution of compound at room temperature. A suitable crystal was selected under a polarizing microscope and mounted on a XtaLAB Synergy, Dualflex, HyPix diffractometer. Data collection was carried out at 100 K. Using Olex2 [1], the structure was solved using Intrinsic Phasing with the SHELXT program [2] and refined using full-matrix least-squares minimization on F2with the SHELXL refinement package [3], All non-hydrogen atoms were refined anisotropically. See FIG. 26.[1] Dolomanov, O.V., Bourhis, L.J., Gildea, RJ, Howard, J.A.K. & Puschmann, H. (2009), J. Appl. Cryst. 42, 339-341.[2] Sheldrick, G.M. (2015). Acta Cryst. A71, 3-8.[3] Sheldrick, G.M. (2015). Acta Cryst. C71, 3-8.

[0547] Although the foregoing subject matter disclosed herein has been described in some detail by way of illustration and example for purposes of clarity of understanding, it is readily apparent to those of ordinary skill in the art in light of the teachings disclosed herein that certain changes and modifications may be made thereto without departing from the spirit or scope of the subject matter disclosed herein.

[0548] Accordingly, the preceding merely illustrates the principles disclosed herein. It will be appreciated that those skilled in the art will be able to devise various arrangements which, although not explicitly described or shown herein, embody the principles disclosed herein and are included within its spirit and scope. Furthermore, all examples and conditional language recited herein are principally intended to aid the reader in understanding the principles disclosed herein and the concepts contributed by the inventors to furthering the art and are to be construed as being without limitation to such specifically recited examples and conditions. Moreover, all statements herein reciting principles, aspects, and embodiments disclosed herein as well as specific examples thereof, are intended to encompass both structural and functional equivalents thereof. Additionally, it is intended that such equivalents include both currently known equivalents and equivalents developed in the future, i.e., any elements developed that perform the same function, regardless ofAttorney Reference: GERN-202WO structure. Moreover, nothing disclosed herein is intended to be dedicated to the public regardless of whether such disclosure is explicitly recited in the claims.

[0549] The scope of the present disclosure, therefore, is not intended to be limited to the exemplary embodiments shown and described herein.

[0550] In the claims, 35 U.S.C. §112(f) is expressly defined as being invoked for a limitation in the claim only when the exact phrase "means for" or the exact phrase "step for" is recited at the beginning of such limitation in the claim; if such exact phrase is not used in a limitation in the claim, then 35 U.S.C. § 112(f) not invoked.

Claims

Attorney Reference: GERN-202WOCLAIMSWhat Is Claimed Is:

1. An isolated enantiomer of Formula (I):or a pharmaceutically acceptable salt thereof, wherein: n is 0 or an integer from 1 to 5;R1and R2are independently selected from H, alkyl, cycloalkyl, heterocyclyl, and substituted versions thereof, wherein one of R1and R2is absent; and each R3is independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof, or provided that there are two adjacent R3groups, the two adjacent R3groups along with the atoms to which they are attached can form a cyclic ring.

2. The isolated enantiomer of claim 1, wherein the isolated enantiomer has the Formula (II):(ID or a pharmaceutically acceptable salt thereof, wherein: n is 0 or an integer from 1 to 4; andA is selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof.

3. The isolated enantiomer of claim 2, wherein n is 1 or 2.Attorney Reference: GERN-202WO4. The isolated enantiomer of claims 2 or 3, wherein R1and R2are independently selected from H and methyl, wherein one of R1and R2is absent.

5. The isolated enantiomer of any one of claims 2-4, wherein each R3is halo.

6. The isolated enantiomer of any one of claims 2-5, wherein:each R4, R5and R6is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof; each R7is independently H or alkyl; and each R8and R9is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof.

7. The isolated enantiomer of claim 6, wherein8. The isolated enantiomer of claim 7, wherein each R4is H.

9. The isolated enantiomer of claims 7 or 8, wherein R5and R6are independently selected from H, alkyl and hydroxyl.

10. The isolated enantiomer of claim 6, wherein A isAttorney Reference: GERN-202WO11. The isolated enantiomer of claim 10, wherein R7is H.

12. The isolated enantiomer of claims 10 or 11, wherein each R8is H.

13. The isolated enantiomer of any one of claims 10-12, wherein R9is carboxy or cyano.

14. The isolated enantiomer of claim 1, wherein the isolated enantiomer has a structure selectedAttorney Reference: GERN-202WO15. The isolated enantiomer of claim 1, having the following structure:Attorney Reference: GERN-202WO16. The isolated enantiomer of claim 1, having the following structure: omer of claim 1, having the following structure:

18. The isolated enantiomer of claim 1, having the following structure:

19. The isolated enantiomer of claim 1, having the following structure:

20. The isolated enantiomer of claim 1, having the following structure:Attorney Reference: GERN-202WO21. The isolated enantiomer of claim 1, having the following structure: iomer of claim 1, having the following structure:

23. The isolated enantiomer of claim 1, having the following structure:

24. The isolated enantiomer of claim 1, having the following structure: iomer of claim 1, having the following structure:Attorney Reference: GERN-202WO26. The isolated enantiomer of claim 1, having the following structure: mer of claim 1, having the following structure:

28. The isolated enantiomer of claim 1, having the following structure:

30. The isolated enantiomer of claim 1, having the following structure:Attorney Reference: GERN-202WO31. The isolated enantiomer of claim 1, having the following structure: omer of claim 1, having the following structure:

33. The isolated enantiomer of claim 1, having the following structure:Attorney Reference: GERN-202WO35. The isolated enantiomer of claim 1, having the following structure:

36. The isolated enantiomer of claim 1, having the following structure:

37. The isolated enantiomer of claim 1, having the following structure: ntiomer of claim 1, having the following structure:

39. The isolated enantiomer of claim 1, having the following structure:Attorney Reference: GERN-202WO40. The isolated enantiomer of claim 1, wherein the isolated enantiomer has a structure selected from the following:Attorney Reference: GERN-202WOAttorney Reference: GERN-202WO41. The isolated enantiomer of claim 1, having the following structure:

42. The isolated enantiomer of claim 1, having the following structure:Attorney Reference: GERN-202WO43. The isolated enantiomer of claim 1, having the following structure: omer of claim 1, having the following structure:

45. The isolated enantiomer of claim 1, having the following structure:

46. The isolated enantiomer of claim 1, having the following structure: ntiomer of claim 1, having the following structure:Attorney Reference: GERN-202WO48. The isolated enantiomer of claim 1, having the following structure:

49. The isolated enantiomer of claim 1, having the following structure:

50. The isolated enantiomer of claim 1, having the following structure:

51. The isolated enantiomer of claim 1, having the following structure:

52. The isolated enantiomer of claim 1, having the following structure:Attorney Reference: GERN-202WO53. The isolated enantiomer of claim 1, having the following structure:

54. The isolated enantiomer of claim 1, having the following structure:

55. The isolated enantiomer of claim 1, having the following structure:

56. The isolated enantiomer of claim 1, having the following structure: tiomer of claim 1, having the following structure:Attorney Reference: GERN-202WO58. The isolated enantiomer of claim 1, having the following structure: omer of claim 1, having the following structure:

60. The isolated enantiomer of claim 1, having the following structure: tiomer of claim 1, having the following structure:

62. The isolated enantiomer of claim 1, having the following structure:Attorney Reference: GERN-202WO63. The isolated enantiomer of claim 1, having the following structure:

64. The isolated enantiomer of claim 1, having the following structure:

65. The isolated enantiomer of claim 1, having the following structure:

66. The isolated enantiomer of claim 1, having the following structure: iomer of claim 1, having the following structure:Attorney Reference: GERN-202WO68. The isolated enantiomer of claim 1, having the following structure: mer of claim 1, having the following structure:

70. The isolated enantiomer of claim 1, having the following structure: mer of claim 1, having the following structure:

72. The isolated enantiomer of claim 1, having the following structure:Attorney Reference: GERN-202WO73. The isolated enantiomer of claim 1, having the following structure:

75. The isolated enantiomer of claim 1, having the following structure: omer of claim 1, having the following structure:

77. The isolated enantiomer of claim 1, having the following structure:Attorney Reference: GERN-202WO78. The isolated enantiomer of claim 1, having the following structure:

79. The isolated enantiomer of claim 1, having the following structure:

80. The isolated enantiomer of claim 1, having the following structure: omer of claim 1, having the following structure:Attorney Reference: GERN-202WO82. The isolated enantiomer of claim 1, having the following structure: mer of claim 1, having the following structure:

84. The isolated enantiomer of claim 1, having the following structure:Attorney Reference: GERN-202WO86. The isolated enantiomer of claim 1, having the following structure:

87. The isolated enantiomer of claim 1, having the following structure:

88. The isolated enantiomer of claim 1, having the following structure:

89. The isolated enantiomer of claim 1, having the following structure:

90. The isolated enantiomer of claim 1, having the following structure:Attorney Reference: GERN-202WO91. The isolated enantiomer of claim 1, having the following structure: ntiomer of claim 1, having the following structure:

93. The isolated enantiomer of claim 1, having the following structure: ntiomer of claim 1, having the following structure:Attorney Reference: GERN-202WO95. The isolated enantiomer of claim 1, having the following structure: tiomer of claim 1, having the following structure:

97. The isolated enantiomer of claim 1, having the following structure: tiomer of claim 1, having the following structure:

99. A pharmaceutical composition comprising at least one isolated enantiomer of any one of claims 1-98, or a pharmaceutically acceptable salt thereof.

100. The pharmaceutical composition of claim 99, further comprising at least one second therapeutic agent.

101. The pharmaceutical composition of claim 100, wherein the at least one second therapeutic agent is imetelstat or imetelstat sodium.Attorney Reference: GERN-202WO102. A method of treating a patient for a disease or condition, the method comprising: administering the pharmaceutical composition of any one of claims 99-101 to the patient.

103. The method of claim 102, wherein the disease or condition is a cancer.

104. The method of claim 103, wherein the cancer is at least one of hematological malignancies and solid tumors.

105. The method of claim 103, wherein the cancer is at least one of acute and chronic leukemias, lymphomas, multiple myeloma, myelodysplastic syndromes (MDS), myeloproliferative neoplasms (MPNs), essential thrombocythemia (ET), polycythemia vera (PV), Chronic Myelogenous Leukemia (CML), myelofibrosis (MF), acute myelogenous leukemia (AML), lung cancers, bladder cancers, liver cancers, colon cancers, adrenal cancers, esophageal cancers, stomach cancers, central nervous system (CNS) cancers, skin cancers, ovarian cancers, cervical cancers, endometrial cancers, renal cancers, prostate cancers, and breast cancers.

106. A method of treating a patient for a disease or condition, the method comprising: administering a therapeutically effective amount of at least one isolated enantiomer of any one of claims 1-98 or a pharmaceutically acceptable salt thereof to the patient.

107. The method of claim 106, wherein the disease or condition is a cancer.

108. The method of claim 107, wherein the cancer is at least one of hematological malignancies and solid tumors.

109. The method of claim 107, wherein the cancer is at least one of acute and chronic leukemias, lymphomas, multiple myeloma, myelodysplastic syndromes (MDS), myeloproliferative neoplasms (MPNs), essential thrombocythemia (ET), polycythemia vera (PV), Chronic Myelogenous Leukemia (CML), myelofibrosis (MF), acute myelogenous leukemia (AML), lung cancers, bladder cancers, liver cancers, colon cancers, adrenal cancers, esophageal cancers, stomach cancers, centralAttorney Reference: GERN-202WO nervous system (CNS) cancers, skin cancers, ovarian cancers, cervical cancers, endometrial cancers, renal cancers, prostate cancers, and breast cancers.

110. A method of treating a patient for a solid tumor comprising: administering a therapeutically effective amount of at least one compound of Formula (I) to the patient:or a pharmaceutically acceptable salt thereof, wherein: n is 0 or an integer from 1 to 5;R1and R2are independently selected from H, alkyl, cycloalkyl, heterocyclyl, and substituted versions thereof, wherein one of R1and R2is absent; and each R3is independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof, or provided that there are two adjacent R3groups, the two adjacent R3groups along with the atoms to which they are attached can form a cyclic ring.

111. The method of claim 110, wherein the at least one compound has the Formula (II):or a pharmaceutically acceptable salt thereof, wherein: n is 0 or an integer from 1 to 4; andAttorney Reference: GERN-202WOA is selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, and substituted versions thereof.

112. The method of claim 111, wherein n is 1 or 2.

113. The method of claims 111 or 112, wherein R1and R2are independently selected from H and methyl, wherein one of R1and R2is absent.

114. The method of any one of claims 111-113, wherein each R3is halo.

115. The method of any one of claims 111-114, wherein:each R4, R5and R6is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof; each R7is independently H or alkyl; and each R8and R9is independently selected from H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, acyl, alkoxy, amino, azido, carbonyl, carboxy, cyano, ether, halo, hydroxy, nitro, thiol, thioether, and substituted versions thereof.

116. The method of claim 115, wherein117. The method of claim 116, wherein each R4is H.Attorney Reference: GERN-202WO118. The method of claims 116 or 117, wherein R5and R6are independently selected from H, alkyl and hydroxyl.

119. The method of claim 115, wherein A is.

120. The method of claim 119, wherein R7is H.

121. The method of claims 119 or 120, wherein each R8is H.

122. The method of any one of claims 119-121, wherein R9is carboxy or cyano.

123. The method of claim 110, wherein the at least one compound has a structure selected from:Attorney Reference: GERN-202WO124. The method of claim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO125. The method of claim 110, wherein the at least one compound has the structure: aim 110, wherein the at least one compound has the structure:

127. The method of claim 110, wherein the at least one compound has the structure:

128. The method of claim 110, wherein the at least one compound has the structure:

129. The method of claim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO130. The method of claim 110, wherein the at least one compound has the structure: aim 110, wherein the at least one compound has the structure:

132. The method of claim 110, wherein the at least one compound has the structure: aim 110, wherein the at least one compound has the structure:

134. The method of claim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO135. The method of claim 110, wherein the at least one compound has the structure: 110, wherein the at least one compound has the structure:

137. The method of claim 110, wherein the at least one compound has the structure:

139. The method of claim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO140. The method of claim 110, wherein the at least one compound has the structure: im 110, wherein the at least one compound has the structure:

142. The method of claim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO144. The method of claim 110, wherein the at least one compound has the structure:

146. The method of claim 110, wherein the at least one compound has the structure: claim 110, wherein the at least one compound has the structure: claim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO149. The method of claim 110, wherein the at least one compound has a structure selected from:Attorney Reference: GERN-202WOAttorney Reference: GERN-202WO150. The method of claim 110, wherein the at least one compound has the structure:

151. The method of claim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO152. The method of claim 110, wherein the at least one compound has the structure: im 110, wherein the at least one compound has the structure:

154. The method of claim 110, wherein the at least one compound has the structure:

155. The method of claim 110, wherein the at least one compound has the structure:

156. The method of claim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO157. The method of claim 110, wherein the at least one compound has the structure:

158. The method of claim 110, wherein the at least one compound has the structure:

159. The method of claim 110, wherein the at least one compound has the structure:

160. The method of claim 110, wherein the at least one compound has the structure:

161. The method of claim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO162. The method of claim 1101, wherein the at least one compound has the structure:

163. The method of claim 110, wherein the at least one compound has the structure:

164. The method of claim 110, wherein the at least one compound has the structure:

165. The method of claim 110, wherein the at least one compound has the structure: aim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO167. The method of claim 110, wherein the at least one compound has the structure: aim 110, wherein the at least one compound has the structure:

169. The method of claim 110, wherein the at least one compound has the structure: im 110, wherein the at least one compound has the structure:

171. The method of claim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO172. The method of claim 110, wherein the at least one compound has the structure:

173. The method of claim 110, wherein the at least one compound has the structure:

174. The method of claim 110, wherein the at least one compound has the structure:

175. The method of claim 110, wherein the at least one compound has the structure: aim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO177. The method of claim 110, wherein the at least one compound has the structure: 110, wherein the at least one compound has the structure:

179. The method of claim 110, wherein the at least one compound has the structure: 110, wherein the at least one compound has the structure:

181. The method of claim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO182. The method of claim 110, wherein the at least one compound has the structure:

184. The method of claim 110, wherein the at least one compound has the structure: im 110, wherein the at least one compound has the structure:

186. The method of claim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO187. The method of claim 110, wherein the at least one compound has the structure:

188. The method of claim 110, wherein the at least one compound has the structure:

189. The method of claim 110, wherein the at least one compound has the structure: im 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO191. The method of claim 110, wherein the at least one compound has the structure: m 110, wherein the at least one compound has the structure:

193. The method of claim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO195. The method of claim 110, wherein the at least one compound has the structure:

197. The method of claim 110, wherein the at least one compound has the structure:

199. The method of claim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO200. The method of claim 110, wherein the at least one compound has the structure: claim 110, wherein the at least one compound has the structure: claim 110, wherein the at least one compound has the structure:

203. The method of claim 110, wherein the at least one compound has the structure:Attorney Reference: GERN-202WO204. The method of claim 110, wherein the at least one compound has the structure: aim 110, wherein the at least one compound has the structure:

206. The method of claim 110, wherein the at least one compound has the structure: aim 110, wherein the at least one compound has the structure:

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