MDM2 targeting protacs

Abstract

Disclosed herein are compounds that may be used to degrade mdm2 that contain one or more rigid linkers in the connection between the mdm2 targeting compound and the degrader. These compound may show improved activity in reducing the mdm2 levels. These compound may be used to treating a disease or disorder such as cancer.

Description

[0001] MDM2 TARGETING PROTACS

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of priority to United States Provisional Application No. 63 / 669,349, filed on July 10, 2024, the entire contents of which is hereby incorporated by reference.

[0004] BACKGROUND

[0005] I. Field

[0006] This disclosure relates to the fields of biology, pharmacology, medicine, and chemistry. In particular, new compounds, compositions, and methods of treatment related to the treatment of cancers are disclosed.

[0007] II. Description of Related Art

[0008] The tumor suppressor p53, a transcription factor, has an essential role in the prevention of human cancer (Levine, 2020). In the absence of cellular stress, the p53 protein is maintained at low levels due to binding to Mdm2, an E3 ubiquitin ligase (Eischen & Lozano, 2014). Mdm2's E3 ligase activity polyubiquitinates p53 resulting in its degradation by the 26S proteasome. p53 is activated by a variety of tumorigenic stresses, which include aberrant oncogene signaling and DNA damage. These stresses activate p53 causing an increase in its transcriptional activity, resulting cell cycle arrest, cellular senescence, and / or apoptosis (Levine, 2020).

[0009] The most well-known function of Mdm2 is as a negative regulator of p53 actions through an auto-regulatory negative feedback loop in which increased p53 levels activate its transcription of Mdm2. Mdm2 then binds p53 blocking its transactivation domain and targets it for proteasomal degradation (Eischen & Lozano, 2014). Since Mdm2 is overproduced in many human cancers (Oliner et al., 2016; Rayburn et al., 2005), targeting its activity is a major focus of drug discovery efforts. Drugs targeting Mdm2, such as RG7112, Idasanutlin, and AMG232 have demonstrated clinical efficacy, and Idasanutlin has progressed into Phase 3 clinical trials (Skalniak et al., 2018; Burgess et al., 2016). However, these drugs have significant undesired toxicities associated with increased levels of Mdm2. Mdm2 inhibitors bind and stabilize the Mdm2 protein by specific binding to the hydrophobic binding pocket of Mdm2. The combination of stabilization of Mdm2 by inhibitor binding and increased p53 levels, which activate Mdm2 transcription, results in significantly increased levels of Mdm2 (Hunziker et al., 2010; Harris & Levine, 2005). Higher Mdm2 levels engage both p53- dependent and p53 -independent functions of Mdm2 (Riley & Lozano, 2012; Eischen & Lozano, 2014; Bouska & Eischen 2009), leading to toxicities (thrombocytopenia and neutropenia), which limits prolonged drug treatment in patients treated with Mdm2 inhibitors (Ray-Coquard et al., 2012). In addition, prolonged treatment with Mdm2 inhibitors leads to resistance caused by p53 mutation (Skalniak et al., 2018; Aziz et al., 2011; Michaelis et al., 2011; Jung et al., 2016).

[0010] Half of all human cancers have inactivated p53 by mutation or deletion (Olivier et al., 2010). To test the requirements of Mdm2 in the absence of p53, Mdm2 was inducibly deleted in primary murine 53 -null lymphoma and sarcoma cells. Mdm2 loss resulted in apoptosis in vitro and in vivo in 5.?-null cancers, with a significantly reduced tumor burden and survival benefit (Feeley et al., 2017). This suggests that Mdm2 may be required for the survival of cancer cells that lack p53. It has been reported that Mdm2 has p53 -independent functions by binding and regulating other proteins, such as p53 family members, p73 and p63, Nbsl in the Mrel l-Rad50-Nbsl DNA break repair complex and other proteins that impact cancer (Riley & Lozano, 2012; Melo & Eischen, 2012; Ongkeko et al., 1999; Zeng et al., 1999; Wu & Leng, 2015). Notably, it was shown that p73 was induced and necessary for mediating the apoptotic effects of Mdm2 loss in the absence of p53 in murine cancer cells (Feeley et al., 2017). The evaluation of human cancer cells that harbor mutant p53 also show that Mdm2 loss induces p73 and apoptosis, indicating this is not a mouse-specific effect. Importantly, p73 has a homologous N-terminal transactivation domain to p53 that binds to Mdm2 in the same site as p53, shown by biophysical studies (Shin et al., 2015), suggesting a remarkably close mimicry of the Mdm2 recognition mechanism among p53 family members. Additionally, unlike p53, p73 is rarely inactivated in human cancers (Dotsch et al., 2010; Venkatanaray an et al., 2016).

[0011] Therefore, there remains a need to develop Mdm2 targeting molecules without the side effects of the known Mdm2 inhibitors and to be able to target p53 -inactivated cancers. SUMMARY

[0012] In some aspects, the present disclosure provides compounds, pharmaceutical compositions, and methods for their use in the treatment of cancers. In some embodiments, the present disclosure relates to Mdm2 degrading molecules such as an Mdm2 targeting PROTAC. In particular, the Mdm2 degrading molecules are described that are able to target both wild-type, mutated, inactivated, and null p53 cancers.

[0013] In another aspect, the present disclosure provides compounds of the formula: wherein:

[0014] Ra, Rb, and Rcare each independently hydrogen, alkylc<i2, or substituted alkylcc ;

[0015] Ri, R2, R2', R3, and R3' are each independently alkylc is, substituted alkylc is, arylccis, or substituted arylc is;

[0016] Y is ((CH2)xO)y, wherein x is 1, 2, or 3 and y is 1, 2, or 3 or Y is taken together with Rato form a heterocycloalkanec<i2 or substituted hctcrocycloalkancc n;

[0017] A is a covalent bond, C(O), alkanediylc<8, or substituted alkancdiylc s; n is 1, 2, or 3;

[0018] R4 is alkylc<8, substituted alkylc<8, arylc<8, substituted arylc<8, aralkylc<8, substituted aralkylc<8, or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group;

[0019] R4' is hydrogen or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkaneccs group;

[0020] Rs is hydroxy, alkoxyc<6, or substituted alkoxyc cl and

[0021] Re and R7 are each independently selected from alkylc<8 or substituted alkylc s; or a pharmaceutically acceptable salt thereof. In some embodiments, the compounds are further defined as: wherein:

[0022] Rb and Rcare each independently hydrogen, alkylc<i2, or substituted alkylc<i2; Ri, R2, R2', R3, and R3' are each independently alkylc<i2, substituted alkylc<i2, arylc<i8, or substituted arylc<is; m is 0, 1, 2, or 3;

[0023] A is a covalent bond, C(O), alkanediylc<8, or substituted alkanediylc<s; n is 1, 2, or 3; R4 is alkylc<8, substituted alkylc<8, arylc<8, substituted arylc<8, aralkylc<8, substituted aralkylc<8, or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group;

[0024] R4' is hydrogen or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group; R5 is hydroxy, alkoxyc<6, or substituted alkoxyc<e; and

[0025] Re and R7 are each independently selected from alkylc<8 or substituted alkylc<s; or a pharmaceutically acceptable salt thereof.

[0026] In some embodiments, the compounds are further defined as: wherein:

[0027] Rb and Rcare each independently hydrogen, alkylc<i2, or substituted alkylc<i2; m is 0, 1, 2, or 3;

[0028] A is a covalent bond, C(O), alkanediylc<8, or substituted alkancdiylc s; n is 1, 2, or 3;

[0029] R4 is alkylc<8, substituted alkylc<8, arylc<8, substituted arylc<8, aralkylc<8, substituted aralkylc<8, or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group;

[0030] R4' is hydrogen or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group;

[0031] Rs is hydroxy, alkoxyc<6, or substituted alkoxyc<e; and

[0032] Re and R7 are each independently selected from alkylc<8 or substituted alkylc<s; or a pharmaceutically acceptable salt thereof.

[0033] In some embodiments, the compounds are further defined as: wherein:

[0034] Rb and Rcare each independently hydrogen, alkylc<i2, or substituted alkylc<i2; m is 0, 1, 2, or 3;

[0035] A is a covalent bond, C(O), alkanediylc<8, or substituted alkancdiylc s; n is 1, 2, or 3;

[0036] R4 is alkylc<8, substituted alkylc<8, arylc<8, substituted arylc<8, aralkylc<8, substituted aralkylc<8, or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group;

[0037] R4' is hydrogen or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group;

[0038] Rs is hydroxy, alkoxyc<6, or substituted alkoxyc<e; and

[0039] Re and R7 are each independently selected from alkylc<8 or substituted alkylc<s; or a pharmaceutically acceptable salt thereof.

[0040] In some embodiments, the compounds are further defined as: wherein:

[0041] Rb and Rcare each independently hydrogen, alkylc<i2, or substituted alkylc<i2; m is 0, 1, 2, or 3;

[0042] A is a covalent bond, C(O), alkanediylc<8, or substituted alkancdiylc s; n is 1, 2, or 3;

[0043] R4 is alkylc<8, substituted alkylc<8, arylc<8, substituted arylc<8, aralkylc<8, substituted aralkylc<8, or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group;

[0044] R4' is hydrogen or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group; and

[0045] Re is alkylc<8 or substituted alkylc<s; or a pharmaceutically acceptable salt thereof.

[0046] In some embodiments, Rais hydrogen. In some embodiments, Y is ((CH2)xO)y, wherein x is 1, 2, or 3 and y is 1, 2, or 3. In some embodiments, x is 1. In other embodiments, x is 2. In some embodiments, y is 1. In other embodiments, Y is taken together with Rato form a heterocycloalkanec<i2 or substituted heterocycloalkanec<i2- In some embodiments, the heterocycloalkanec<i2 is a pyrrolidine, a piperidine, an azepane, an azocane or an alkyl substituted version of any of these compounds.

[0047] In some embodiments, Ri is substituted arylc<is such as 2-ethoxy-4-t-butylphenyl. In some embodiments, R2 and R2' are different. In some embodiments, the carbon atom to which R2 and R2' are attached is in the R configuration. In other embodiments, the carbon atom to which R2 and R2' are attached is in the S configuration. In some embodiments, R2 is substituted arylc<i8 such as 4-chlorophenyl. In some embodiments, R2' is alkylc<8 such as methyl. In some embodiments, R3 and R3' are different. In some embodiments, the carbon atom to which R3 and R3' are attached is in the R configuration. In other embodiments, the carbon atom to which R3 and R3' are attached is in the S configuration. In some embodiments, R3 is substituted arylc<is such as 4-chlorophenyl. In some embodiments, R3' is alkylc<8 such as methyl.

[0048] In some embodiments, A is a covalent bond. In other embodiments, A is C(O). In other embodiments, A is alkanediylc<8 or substituted alkanediylc<8. In some embodiments, A is alkanediylc<8 such as methylene or ethylene. In some embodiments, m is 1. In other embodiments, m is 2. In other embodiments, m is 3. In some embodiments, n is 1. In other embodiments, n is 2.

[0049] In some embodiments, R4 and R4' are different. In some embodiments, the carbon atom to which R4 and R4' are attached is in the R configuration. In other embodiments, the carbon atom to which R4 and R4' are attached is in the S configuration. In some embodiments, R4' is hydrogen. In other embodiments, R4 is taken together with R4' to form a cycloalkanec<8 or substituted cycloalkanec<8 group. In some embodiments, R4 is taken together with R4' to form a cycloalkanec<8 group. In some embodiments, the cycloalkanec<8 group is a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, or a cyclohexyl group. In other embodiments, R4 is alkylc<8 or substituted alkylc<8. In some embodiments, R4 is alkylc<8 such as t-butyl. In other embodiments, R4 is aralkylc<8 or substituted aralkylc<8- In some embodiments, R4 is aralkylc<8 such as benzyl. In some embodiments, R4 is arylc<8 or substituted arylc<8- In some embodiments, R4 is arylc<8 such as phenyl.

[0050] In some embodiments, Rb is hydrogen. In some embodiments, Rcis hydrogen. In some embodiments, the carbon atom to which R5 is attached is in the R configuration. In other embodiments, the carbon atom to which R5 is attached is in the S configuration. In some embodiments, R5 is hydroxy. In some embodiments, the carbon atom to which Re is attached is in the R configuration. In other embodiments, the carbon atom to which Re is attached is in the S configuration. In some embodiments, Re is hydrogen. In other embodiments, Re is alkylc<8 such as methyl. In some embodiments, R7 is alkylc<8 such as methyl. In some embodiments, the compound is further defined as:

[0051] or a pharmaceutically acceptable salt thereof.

[0052] In still another aspect, the present disclosure provides pharmaceutical composition comprising:

[0053] (A) a compound described herein; and

[0054] (B) a pharmaceutically acceptable excipient.

[0055] In some embodiments, the pharmaceutical compositions are formulated for oral administration or administration via injection. In some embodiments, the pharmaceutical compositions are formulated as a unit dose.

[0056] In still another aspect, the present disclosure provides methods of treating a disease or disorder in a patient in need thereof comprising administering to the patient in need thereof a therapeutically effective amount of a compound or composition described herein. In some embodiments, the disease or disorder is cancer. In some embodiments, the cancer is a carcinoma, sarcoma, lymphoma, leukemia, melanoma, mesothelioma, multiple myeloma, or seminoma. In some embodiments, the cancer is of the bladder, blood, bone, brain, breast, central nervous system, cervix, colon, endometrium, esophagus, gall bladder, gastrointestinal tract, genitalia, genitourinary tract, head, kidney, larynx, liver, lung, muscle tissue, neck, oral or nasal mucosa, ovary, pancreas, prostate, skin, spleen, small intestine, large intestine, stomach, testicle, or thyroid.

[0057] In some embodiments, the cancer comprises a mutated p53 gene. In some embodiments, the cancer comprises a deleted p53 gene. In some embodiments, the cancer is breast cancer such as triple negative breast cancer. In other embodiments, the cancer is ovarian cancer. In other embodiments, the cancer is Burkitt lymphoma. In other embodiments, the cancer is diffuse large B cell lymphoma. In some embodiments, the diffuse large B cell lymphoma is subtype ABC. In other embodiments, the diffuse large B cell lymphoma is subtype GCB. In other embodiments, the cancer is a B-cell leukemia such as B-cell acute lymphocytic leukemia. In other embodiments, the cancer is a T-cell leukemia such as T-cell acute lymphocytic leukemia. In some embodiments, the cancer is a lung cancer such as lung adenocarcinoma. In other embodiments, the cancer is head and neck cancer. In other embodiments, the cancer is a sarcoma.

[0058] In some embodiments, the methods further comprise administering a second therapy. In some embodiments, the second therapy is surgery, a second chemotherapeutic, radiotherapy, or immunotherapy. In some embodiments, the compound is administered once. In other embodiments, the compound is administered two or more times.

[0059] In another aspect, the present disclosure provides methods of inhibiting cell replication by modulating mdm2 comprising contacting the cell with a compound or composition described herein.

[0060] In still yet another aspect, the present disclosure provides methods of modulating the activity of mdm2 in a cell comprising contacting the cell with a compound or composition described herein.

[0061] In some embodiments, the method is performed in vitro. In some embodiments, the method is performed in vivo. In some embodiments, the method is performed ex vivo.

[0062] Other objects, features and advantages of the present disclosure will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating specific embodiments of the disclosure, are given by way of illustration only, since various changes and modifications within the spirit and scope of the disclosure will become apparent to those skilled in the art from this detailed description. Note that simply because a particular compound is ascribed to one particular generic formula doesn’t mean that it cannot also belong to another generic formula.

[0063] BRIEF DESCRIPTION OF THE DRAWINGS

[0064] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present disclosure. The disclosure may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein.

[0065] FIG. 1 shows the quantification of the Mdm2 protein levels based on Western blot analysis after 16-hour treatment with indicated compounds in NanoLuc-Mdm2 expressing 293T cells relative to control at treatment concentrations of 0, 8, 40, 200, and 1000 nM.

[0066] FIG. 2 shows the YX-02-030 and IK-01-189 activity in reducing MDM2 protein levels in the MDM2-NanoLuc expressing 293T cells with GAPDH acting as a loading control as measured by Western blot analysis.

[0067] FIG. 3 shows the MKM-01-143 and MKM-01-219 activity in reducing MDM2 protein levels in the MDM2-NanoLuc expressing 293T cells with GAPDH acting as a loading control as measured by Western blot analysis. FIG. 4 shows the MKM-01-153 and MKM-01-217 activity in reducing MDM2 protein levels in the MDM2-NanoLuc expressing 293T cells with GAPDH acting as a loading control as measured by Western blot analysis.

[0068] DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0069] Provided herein are compounds that may be used to degrade Mdm2 and thus may be used in the treatment of cancers including cancers associated with overexpression of Mdm2 and / or cancers which may be treated by modulating the activity of p53. These compounds may show improved tolerability and stability compared to traditional inhibitors of Mdm2. These and more details will be described below.

[0070] I. Compounds and Formulations Thereof

[0071] A. Compounds of the Present Disclosure

[0072] The compounds of the present disclosure are shown, for example, in the summary section above and in Table 1 and the claims below. They may be made using the synthetic methods outlined in the Examples section. These methods can be further modified and optimized using the principles and techniques of organic chemistry as applied by a person skilled in the art. Such principles and techniques are taught, for example, in Smith, March ’s Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, (2013), which is incorporated by reference herein. In addition, the synthetic methods may be further modified and optimized for preparative, pilot- or large-scale production, either batch or continuous, using the principles and techniques of process chemistry as applied by a person skilled in the art. Such principles and techniques are taught, for example, in Anderson, Practical Process Research & Development - A Guide for Organic Chemists (2012), which is incorporated by reference herein.

[0073] Table 1: Compounds of the Present Disclosure

[0074]

[0075]

[0076] All the compounds of the present disclosure may in some embodiments be used for the prevention and treatment of one or more diseases or disorders discussed herein or otherwise. In some embodiments, one or more of the compounds characterized or exemplified herein as an intermediate, a metabolite, and / or prodrug, may nevertheless also be useful for the prevention and treatment of one or more diseases or disorders. As such unless explicitly stated to the contrary, all the compounds of the present disclosure are deemed “active compounds” and “therapeutic compounds” that are contemplated for use as active pharmaceutical ingredients (APIs). Actual suitability for human or veterinary use is typically determined using a combination of clinical trial protocols and regulatory procedures, such as those administered by the Food and Drug Administration (FDA). In the United States, the FDA is responsible for protecting the public health by assuring the safety, effectiveness, quality, and security of human and veterinary drugs, vaccines and other biological products, and medical devices. In some embodiments, the compounds of the present disclosure have the advantage that they may be more efficacious than, be less toxic than, be longer acting than, be more potent than, produce fewer side effects than, be more easily absorbed than, more metabolically stable than, more lipophilic than, more hydrophilic than, and / or have a better pharmacokinetic profile (e.g., higher oral bioavailability and / or lower clearance) than, and / or have other useful pharmacological, physical, or chemical properties over, compounds known in the prior art, whether for use in the indications stated herein or otherwise.

[0077] Compounds of the present disclosure may contain one or more asymmetrically- substituted carbon or nitrogen atom and may be isolated in optically active or racemic form. Thus, all chiral, diastereomeric, racemic form, epimeric form, and all geometric isomeric forms of a chemical formula are intended, unless the specific stereochemistry or isomeric form is specifically indicated. Compounds may occur as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. In some embodiments, a single diastereomer is obtained. The chiral centers of the compounds of the present disclosure can have the S or the R configuration. In some embodiments, the present compounds may contain two or more atoms which have a defined stereochemical orientation.

[0078] Chemical formulas used to represent compounds of the present disclosure will typically only show one of possibly several different tautomers. For example, many types of ketone groups are known to exist in equilibrium with corresponding enol groups. Similarly, many types of imine groups exist in equilibrium with enamine groups. Regardless of which tautomer is depicted for a given compound, and regardless of which one is most prevalent, all tautomers of a given chemical formula are intended.

[0079] In addition, atoms making up the compounds of the present disclosure are intended to include all isotopic forms of such atoms. Isotopes, as used herein, include those atoms having the same atomic number but different mass numbers. By way of general example and without limitation, isotopes of hydrogen include tritium and deuterium, and isotopes of carbon include13C and14C.

[0080] In some embodiments, compounds of the present disclosure exist in salt or non-salt form. With regard to the salt form(s), in some embodiments the particular anion or cation forming a part of any salt form of a compound provided herein is not critical, so long as the salt, as a whole, is pharmacologically acceptable. Additional examples of pharmaceutically acceptable salts and their methods of preparation and use are presented in Handbook of Pharmaceutical Salts: Properties, and Use (2002), which is incorporated herein by reference.

[0081] B. Pharmaceutical Formulations and Routes of Administration

[0082] In another aspect, for administration to a patient in need of such treatment, pharmaceutical formulations (also referred to as a pharmaceutical preparations, pharmaceutical compositions, pharmaceutical products, medicinal products, medicines, medications, or medicaments) comprise a therapeutically effective amount of a compound disclosed herein formulated with one or more excipients and / or drug carriers appropriate to the indicated route of administration. In some embodiments, the compounds disclosed herein are formulated in a manner amenable for the treatment of human and / or veterinary patients. In some embodiments, formulation comprises admixing or combining one or more of the compounds disclosed herein with one or more of the following excipients: lactose, sucrose, starch powder, cellulose esters of alkanoic acids, cellulose alkyl esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids, gelatin, acacia, sodium alginate, polyvinylpyrrolidone, and / or polyvinyl alcohol. In some embodiments, e.g., for oral administration, the pharmaceutical formulation may be tableted or encapsulated. In some embodiments, the compounds may be dissolved or slurried in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, and / or various buffers. In some embodiments, the pharmaceutical formulations may be subjected to pharmaceutical operations, such as sterilization, and / or may contain drug carriers and / or excipients such as preservatives, stabilizers, wetting agents, emulsifiers, encapsulating agents such as lipids, dendrimers, polymers, proteins such as albumin, nucleic acids, and buffers.

[0083] Pharmaceutical formulations may be administered by a variety of methods, e.g., orally or by injection (e.g. subcutaneous, intravenous, and intraperitoneal). Depending on the route of administration, the compounds disclosed herein may be coated in a material to protect the compound from the action of acids and other natural conditions which may inactivate the compound. To administer the active compound by other than parenteral administration, it may be necessary to coat the compound with, or co-administer the compound with, a material to prevent its inactivation. In some embodiments, the active compound may be administered to a patient in an appropriate carrier, for example, liposomes, or a diluent. Pharmaceutically acceptable diluents include saline and aqueous buffer solutions. Liposomes include water-in- oil-in-water CGF emulsions as well as conventional liposomes.

[0084] The compounds disclosed herein may also be administered parenterally, intraperitoneally, intraspinally, or intracerebrally. Dispersions can be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms.

[0085] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (such as, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, sodium chloride, or polyalcohols such as mannitol and sorbitol, in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate or gelatin.

[0086] The compounds disclosed herein can be administered orally, for example, with an inert diluent or an assimilable edible carrier. The compounds and other ingredients may also be enclosed in a hard or soft-shell gelatin capsule, compressed into tablets, or incorporated directly into the patient’s diet. For oral therapeutic administration, the compounds disclosed herein may be incorporated with excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. The percentage of the therapeutic compound in the compositions and preparations may, of course, be varied. The amount of the therapeutic compound in such pharmaceutical formulations is such that a suitable dosage will be obtained. The therapeutic compound may also be administered topically to the skin, eye, ear, or mucosal membranes. Administration of the therapeutic compound topically may include formulations of the compounds as a topical solution, lotion, cream, ointment, gel, foam, transdermal patch, or tincture. When the therapeutic compound is formulated for topical administration, the compound may be combined with one or more agents that increase the permeability of the compound through the tissue to which it is administered. In other embodiments, it is contemplated that the topical administration is administered to the eye. Such administration may be applied to the surface of the cornea, conjunctiva, or sclera. Without wishing to be bound by any theory, it is believed that administration to the surface of the eye allows the therapeutic compound to reach the posterior portion of the eye. Ophthalmic topical administration can be formulated as a solution, suspension, ointment, gel, or emulsion. Finally, topical administration may also include administration to the mucosa membranes such as the inside of the mouth. Such administration can be directly to a particular location within the mucosal membrane such as a tooth, a sore, or an ulcer. Alternatively, if local delivery to the lungs is desired the therapeutic compound may be administered by inhalation in a dry -powder or aerosol formulation.

[0087] In some embodiments, it may be advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the patients to be treated; each unit containing a predetermined quantity of therapeutic compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. In some embodiments, the specification for the dosage unit forms of the disclosure are dictated by and directly dependent on (a) the unique characteristics of the therapeutic compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such a therapeutic compound for the treatment of a selected condition in a patient. In some embodiments, active compounds are administered at a therapeutically effective dosage sufficient to treat a condition associated with a condition in a patient. For example, the efficacy of a compound can be evaluated in an animal model system that may be predictive of efficacy in treating the disease in a human or another animal.

[0088] In some embodiments, the effective dose range for the therapeutic compound can be extrapolated from effective doses determined in animal studies for a variety of different animals. In some embodiments, the human equivalent dose (HED) in mg / kg can be calculated in accordance with the following formula (see, e.g., Reagan-Shaw et al., FASEB J., 22(3):659- 661, 2008, which is incorporated herein by reference):

[0089] HED (mg / kg) = Animal dose (mg / kg) x (Animal Km / Hurnan Km)

[0090] Use of the Kmfactors in conversion results in HED values based on body surface area (BSA) rather than only on body mass. Kmvalues for humans and various animals are well known. For example, the Kmfor an average 60 kg human (with a BSA of 1.6 m2) is 37, whereas a 20 kg child (BSA 0.8 m2) would have a Kmof 25. Kmfor some relevant animal models are also well known, including: mice Kmof 3 (given a weight of 0.02 kg and BSA of 0.007); hamster Kmof 5 (given a weight of 0.08 kg and BSA of 0.02); rat Kmof 6 (given a weight of 0.15 kg and BSA of 0.025) and monkey Kmof 12 (given a weight of 3 kg and BSA of 0.24).

[0091] Precise amounts of the therapeutic composition depend on the judgment of the practitioner and are specific to each individual. Nonetheless, a calculated HED dose provides a general guide. Other factors affecting the dose include the physical and clinical state of the patient, the route of administration, the intended goal of treatment and the potency, stability and toxicity of the particular therapeutic formulation.

[0092] The actual dosage amount of a compound of the present disclosure or composition comprising a compound of the present disclosure administered to a patient may be determined by physical and physiological factors such as type of animal treated, age, sex, body weight, severity of condition, the type of disease being treated, previous or concurrent therapeutic interventions, idiopathy of the patient and on the route of administration. These factors may be determined by a skilled artisan. The practitioner responsible for administration will typically determine the concentration of active ingredient(s) in a composition and appropriate dose(s) for the individual patient. The dosage may be adjusted by the individual physician in the event of any complication.

[0093] In some embodiments, the therapeutically effective amount typically will vary from about 0.001 mg / kg to about 1000 mg / kg, from about 0.01 mg / kg to about 750 mg / kg, from about 100 mg / kg to about 500 mg / kg, from about 1 mg / kg to about 250 mg / kg, from about 10 mg / kg to about 150 mg / kg in one or more dose administrations daily, for one or several days (depending of course of the mode of administration and the factors discussed above). Other suitable dose ranges include 1 mg to 10,000 mg per day, 100 mg to 10,000 mg per day, 500 mg to 10,000 mg per day, and 500 mg to 1,000 mg per day. In some embodiments, the amount is less than 10,000 mg per day with a range of 750 mg to 9,000 mg per day.

[0094] In some embodiments, the amount of the active compound in the pharmaceutical formulation is from about 2 to about 75 weight percent. In some of these embodiments, the amount is from about 25 to about 60 weight percent.

[0095] Single or multiple doses of the agents are contemplated. Desired time intervals for delivery of multiple doses can be determined by one of ordinary skill in the art employing no more than routine experimentation. As an example, patients may be administered two doses daily at approximately 12-hour intervals. In some embodiments, the agent is administered once a day.

[0096] The agent(s) may be administered on a routine schedule. As used herein a routine schedule refers to a predetermined designated period of time. The routine schedule may encompass periods of time which are identical, or which differ in length, as long as the schedule is predetermined. For instance, the routine schedule may involve administration twice a day, every day, every two days, every three days, every four days, every five days, every six days, a weekly basis, a monthly basis or any set number of days or weeks there-between. Alternatively, the predetermined routine schedule may involve administration on a twice daily basis for the first week, followed by a daily basis for several months, etc. In other embodiments, the disclosure provides that the agent(s) may be taken orally and that the timing of which is or is not dependent upon food intake. Thus, for example, the agent can be taken every morning and / or every evening, regardless of when the patient has eaten or will eat.

[0097] II. Methods of Treatment and Combination Therapies

[0098] A. Methods of Treatment

[0099] In particular the compositions that may be used in treating a disease or disorder in a subject (e.g., a human subject) are disclosed herein. The compositions described above are preferably administered to a mammal (e.g., rodent, human, non-human primates, canine, bovine, ovine, equine, feline, etc.) in an effective amount, that is, an amount capable of producing a desirable result in a treated subject (e.g. , slowing, stopping, reducing or eliminating one or more symptoms or underlying causes of disease). Toxicity and therapeutic efficacy of the compositions utilized in methods of the disclosure can be determined by standard pharmaceutical procedures. As is well known in the medical and veterinary arts, dosage for any one animal depends on many factors, including the subject's size, body surface area, body weight, age, the particular composition to be administered, time and route of administration, general health, the clinical symptoms and other drugs being administered concurrently. In some embodiments, the amount of the compounds used is calculated to be from about 0.01 mg to about 10,000 mg / day. In some embodiments, the amount is from about 1 mg to about 1,000 mg / day. In some embodiments, the compounds may be administered for 1 day to 20 days. In further embodiments, it is contemplated that the compounds may be administered for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, or 20 days, or any range derivable therein. In some embodiments, the compounds may be administered for between 3 and 5 days, inclusive. In some embodiments, the compounds may be administered once. It is also contemplated that in some embodiments, the compounds disclosed herein may be administered two or more times. In some embodiments, these dosings may be reduced or increased based upon the biological factors of a particular patient such as increased or decreased metabolic breakdown of the drug or decreased uptake by the digestive tract if administered orally. Additionally, the compounds may be more efficacious and thus a smaller dose is required to achieve a similar effect. Such a dose is typically administered once a day for a few weeks or until sufficient achieve clinical benefit.

[0100] The therapeutic methods of the disclosure (which include prophylactic treatment) in general include administration of a therapeutically effective amount of the compositions described herein to a subject in need thereof, including a mammal, particularly a human. Such treatment will be suitably administered to subjects, particularly humans, suffering from, having, susceptible to, or at risk for a disease, disorder, or symptom thereof. Determination of those subjects "at risk" can be made by any objective or subjective determination by a diagnostic test or opinion of a subject or health care provider (e.g., genetic test, enzyme or protein marker, family history, and the like).

[0101] B. Hyperproliferative Diseases

[0102] While hyperproliferative diseases can be associated with any medical disorder that causes a cell to begin to reproduce uncontrollably, the prototypical example is cancer. One of the key elements of cancer is that the normal apoptotic cycle of the cell is interrupted and thus agents that lead to apoptosis of the cell are important therapeutic agents for treating these diseases. As such, the Mdm2 degraders described in this disclosure may be effective in treating cancers. In particular, the Mdm2 degraders may be used to treat one or more cancers that expresses a mutant p53 gene or that have deleted or truncated the p53 gene. The p53 gene may be mutated in such a way that it results in decreased expression, decreased activity, or is nonfunctional. Similarly, the p53 gene may also be the wild type gene. In another embodiments, the p53 gene in the cancer may be null. In particular, the present disclosure relates to the treatment of one or more cancers such as breast cancer including triple negative breast cancer (TNBC). The present disclosure also relates to the treatment of lymphoma such as Burkitt lymphoma, Diffuse large B cell lymphoma, and T cell lymphoma. The present disclosure also relates to the treatment of leukemia such as B cell acute lymphoblastic leukemia (B-ALL) or T cell acute lymphoblastic leukemia (T-ALL). The present disclosure also relates to the treatment of lung cancer such as lung adenocarcinoma. The present disclosure also relates to the treatment of ovarian cancer, head and neck cancer, or sarcoma.

[0103] Cancer cells that may be treated with the compounds according to the embodiments include but are not limited to cells from the bladder, blood, bone, bone marrow, brain, breast, colon, esophagus, gastrointestine, gum, head, kidney, liver, lung, nasopharynx, neck, ovary, prostate, skin, stomach, pancreas, testis, tongue, cervix, or uterus. In addition, the cancer may specifically be of the following histological type, though it is not limited to these: neoplasm, malignant; carcinoma; carcinoma, undifferentiated; giant and spindle cell carcinoma; small cell carcinoma; papillary carcinoma; squamous cell carcinoma; lymphoepithelial carcinoma; basal cell carcinoma; pilomatrix carcinoma; transitional cell carcinoma; papillary transitional cell carcinoma; adenocarcinoma; gastrinoma, malignant; cholangiocarcinoma; hepatocellular carcinoma; combined hepatocellular carcinoma and cholangiocarcinoma; trabecular adenocarcinoma; adenoid cystic carcinoma; adenocarcinoma in adenomatous polyp; adenocarcinoma, familial polyposis coli; solid carcinoma; carcinoid tumor, malignant; branchiolo-alveolar adenocarcinoma; papillary adenocarcinoma; chromophobe carcinoma; acidophil carcinoma; oxyphilic adenocarcinoma; basophil carcinoma; clear cell adenocarcinoma; granular cell carcinoma; follicular adenocarcinoma; papillary and follicular adenocarcinoma; nonencapsulating sclerosing carcinoma; adrenal cortical carcinoma; endometroid carcinoma; skin appendage carcinoma; apocrine adenocarcinoma; sebaceous adenocarcinoma; ceruminous adenocarcinoma; mucoepidermoid carcinoma; cystadenocarcinoma; papillary cystadenocarcinoma; papillary serous cystadenocarcinoma; mucinous cystadenocarcinoma; mucinous adenocarcinoma; signet ring cell carcinoma; infiltrating duct carcinoma; medullary carcinoma; lobular carcinoma; inflammatory carcinoma; paget's disease, mammary; acinar cell carcinoma; adenosquamous carcinoma; adenocarcinoma w / squamous metaplasia; thymoma, malignant; ovarian stromal tumor, malignant; thecoma, malignant; granulosa cell tumor, malignant; androblastoma, malignant; sertoli cell carcinoma; leydig cell tumor, malignant; lipid cell tumor, malignant; paraganglioma, malignant; extramammary paraganglioma, malignant; pheochromocytoma; glomangiosarcoma; malignant melanoma; amelanotic melanoma; superficial spreading melanoma; malig melanoma in giant pigmented nevus; epithelioid cell melanoma; blue nevus, malignant; sarcoma; fibrosarcoma; fibrous histiocytoma, malignant; myxosarcoma; liposarcoma; leiomyosarcoma; rhabdomyosarcoma; embryonal rhabdomyosarcoma; alveolar rhabdomyosarcoma; stromal sarcoma; mixed tumor, malignant; mullerian mixed tumor; nephroblastoma; hepatoblastoma; carcinosarcoma; mesenchymoma, malignant; brenner tumor, malignant; phyllodes tumor, malignant; synovial sarcoma; mesothelioma, malignant; dysgerminoma; embryonal carcinoma; teratoma, malignant; struma ovarii, malignant; choriocarcinoma; mesonephroma, malignant; hemangiosarcoma; hemangioendothelioma, malignant; kaposi's sarcoma; hemangiopericytoma, malignant; lymphangiosarcoma; osteosarcoma; juxtacortical osteosarcoma; chondrosarcoma; chondroblastoma, malignant; mesenchymal chondrosarcoma; giant cell tumor of bone; ewing's sarcoma; odontogenic tumor, malignant; ameloblastic odontosarcoma; ameloblastoma, malignant; ameloblastic fibrosarcoma; pinealoma, malignant; chordoma; glioma, malignant; ependymoma; astrocytoma; protoplasmic astrocytoma; fibrillary astrocytoma; astroblastoma; glioblastoma; oligodendroglioma; oligodendroblastoma; primitive neuroectodermal; cerebellar sarcoma; ganglioneuroblastoma; neuroblastoma; retinoblastoma; olfactory neurogenic tumor; meningioma, malignant; neurofibrosarcoma; neurilemmoma, malignant; granular cell tumor, malignant; malignant lymphoma; Hodgkin's disease; Hodgkin's; paragranuloma; malignant lymphoma, small lymphocytic; malignant lymphoma, large cell, diffuse; malignant lymphoma, follicular; mycosis fungoides; other specified non-Hodgkin's lymphomas; malignant histiocytosis; multiple myeloma; mast cell sarcoma; immunoproliferative small intestinal disease; leukemia; lymphoid leukemia; plasma cell leukemia; erythroleukemia; lymphosarcoma cell leukemia; myeloid leukemia; basophilic leukemia; eosinophilic leukemia; monocytic leukemia; mast cell leukemia; megakaryoblastic leukemia; myeloid sarcoma; and hairy cell leukemia. In certain aspects, the tumor may comprise an osteosarcoma, angiosarcoma, rhabdosarcoma, leiomyosarcoma, Ewing sarcoma, glioblastoma, neuroblastoma, or leukemia. C. Combination Therapies

[0104] In addition to being used as a monotherapy, the compounds of the present invention may also find use in combination therapies. Effective combination therapy may be achieved with a single composition or pharmacological formulation that includes both agents, or with two distinct compositions or formulations, administered at the same time, wherein one composition includes a compound of this invention, and the other includes the second agent(s). Alternatively, the therapy may precede or follow the other agent treatment by intervals ranging from minutes to months.

[0105] To treat diseases or disorders using the methods and compositions of the present disclosure, one would generally contact a cell or a subject with a compound and at least one other therapy. These therapies would be provided in a combined amount effective to achieve a reduction in one or more disease parameter. This process may involve contacting the cells / subjects with both agents / therapies at the same time, e.g., using a single composition or pharmacological formulation that includes both agents, or by contacting the cell / subject with two distinct compositions or formulations, at the same time, wherein one composition includes the compound and the other includes the other agent. In some embodiments, the compounds of the present disclosure or any therapies used in conjunction with the compounds of the present disclosure may be administered in a less than therapeutically effective dose when used either alone or in combination.

[0106] Non-limiting examples of such combination therapy include combination of one or more compounds of the invention with another pro-inflammatory agent, an immunosuppressant agent, a chemotherapeutic agent, radiation therapy, an antidepressant, an antipsychotic agent, an anticonvulsant, a mood stabilizer, an anti-infective agent, an antihypertensive agent, a cholesterol-lowering agent or other modulator of blood lipids, an agent for promoting weight loss, an antithrombotic agent, an agent for treating or preventing cardiovascular events such as myocardial infarction or stroke, an antidiabetic agent, an agent for reducing transplant rejection or graft-versus-host disease, an anti-arthritic agent, an analgesic agent, an anti-asthmatic agent or other treatment for respiratory diseases, or an agent for treatment or prevention of skin disorders. Compounds of the invention may be combined with agents designed to improve a patient’s immune response to cancer, including (but not limited to) cancer vaccines. See Lu et al. (2011), which is incorporated herein by reference. Cancer therapies also include a variety of combination therapies with both chemical and radiation based treatments. Combination chemotherapies include, for example, cisplatin (CDDP), carboplatin, procarbazine, mechlorethamine, cyclophosphamide, camptothecin, ifosfamide, melphalan, chlorambucil, busulfan, nitrosurea, dactinomycin, daunorubicin, doxorubicin, bleomycin, plicomycin, mitomycin, etoposide (VP16), tamoxifen, raloxifene, estrogen receptor binding agents, taxol, gemcitabien, navelbine, farnesyl-protein transferase inhibitors, transplatinum, 5 -fluorouracil, vincristine, vinblastine and methotrexate, Temazolomide (an aqueous form of DTIC), or any analog or derivative variant of the foregoing. The combination of chemotherapy with biological therapy is known as biochemotherapy. The present invention contemplates any chemotherapeutic agent that may be employed or known in the art for treating or preventing cancers.

[0107] Other factors that cause DNA damage and have been used extensively include what are commonly known as y-rays, X-rays, and / or the directed delivery of radioisotopes to tumor cells. Other forms of DNA damaging factors are also contemplated such as microwaves and UV- irradiation. It is most likely that all of these factors effect a broad range of damage on DNA, on the precursors of DNA, on the replication and repair of DNA, and on the assembly and maintenance of chromosomes. Dosage ranges for X-rays range from daily doses of 50 to 200 roentgens for prolonged periods of time (3 to 4 wk), to single doses of 2000 to 6000 roentgens. Dosage ranges for radioisotopes vary widely, and depend on the half-life of the isotope, the strength and type of radiation emitted, and the uptake by the neoplastic cells.

[0108] The terms “contacted” and “exposed,” when applied to a cell, are used herein to describe the process by which a therapeutic agent and a chemotherapeutic or radiotherapeutic agent are delivered to a target cell or are placed in direct juxtaposition with the target cell. To achieve cell killing or stasis, both agents are delivered to a cell in a combined amount effective to kill the cell or prevent it from dividing.

[0109] It is also conceivable that more than one administration of either the compound or the other therapy will be desired. Various combinations may be employed, where a compound of the present disclosure is “A,” and the other therapy is “B,” as exemplified below:

[0110] A / B / A B / A / B B / B / A A / A / B B / A / A A / B / B B / B / B / A B / B / A / B A / A / B / B A / B / A / B A / B / B / A B / B / A / A B / A / B / A B / A / A / B B / B / B / A A / A / A / B B / A / A / A A / B / A / A A / A / B / A A / B / B / B B / A / B / B B / B / A / B Other combinations are also contemplated.

[0111] III. Chemical Definitions

[0112] When used in the context of a chemical group: “hydrogen” means -H; “hydroxy” means -0H; ‘ ‘oxo” means =0; “carbonyl” means -C(=0)-; “carboxy” means -C(=0)0H (also written as -C00H or -CO2H); “halo” means independently -F, -Cl, -Br or -I; “amino” means -NH2; “hydroxyamino” means -NH0H; “nitro” means -NO2; imino means =NH; “cyano” means -CN; “isocyanyl” means -N=C=0; “azido” means -N3; in a monovalent context “phosphate” means -0P(0)(0H)2 or a deprotonated form thereof; in a divalent context “phosphate” means -0P(0)(0H)0- or a deprotonated form thereof; “mercapto” means -SH; and “thio” means =S; “thiocarbonyl” mens -C(=S)-; “sulfonyl” means -S(0)2~; and “sulfinyl” means -S(0)-.

[0113] In the context of chemical formulas, the symbol means a single bond, “=” means a double bond, and “=” means triple bond. The symbol “ - ” represents an optional bond, which if present is either single or double. The symbol “ - ” represents a single bond or a double bond. Thus, the formula covers, for example, 0,0,0,0and And it is understood that no one such ring atom forms part of more than one double bond. Furthermore, it is noted that the covalent bond symbol when connecting one or two stereogenic atoms, does not indicate any preferred stereochemistry. Instead, it covers all stereoisomers as well as mixtures thereof. The symbol “ » AA ”, when drawn perpendicularly for methyl) indicates a point of attachment of the group. It is noted that the point of attachment is typically only identified in this manner for larger groups in order to assist the reader in unambiguously identifying a point of attachment. The symbol means a single bond where the group attached to the thick end of the wedge is “out of the page.” The symbol “ "'Hll ” means a single bond where the group attached to the thick end of the wedge is “into the page”. The symbol “ » AA. ” means a single bond where the geometry around a double bond (e.g., either E or Z) is undefined. Both options, as well as combinations thereof are therefore intended. Any undefined valency on an atom of a structure shown in this application implicitly represents a hydrogen atom bonded to that atom. A bold dot on a carbon atom indicates that the hydrogen attached to that carbon is oriented out of the plane of the paper.

[0114] When a variable is depicted as a “floating group” on a ring system, for example, the group “R” in the formula: then the variable may replace any hydrogen atom attached to any of the ring atoms, including a depicted, implied, or expressly defined hydrogen, so long as a stable structure is formed. When a variable is depicted as a “floating group” on a fused ring system, as for example the group “R” in the formula: then the variable may replace any hydrogen attached to any of the ring atoms of either of the fused rings unless specified otherwise. Replaceable hydrogens include depicted hydrogens (e.g., the hydrogen attached to the nitrogen in the formula above), implied hydrogens e.g., a hydrogen of the formula above that is not shown but understood to be present), expressly defined hydrogens, and optional hydrogens whose presence depends on the identity of a ring atom e.g., a hydrogen attached to group X, when X equals -CH-), so long as a stable structure is formed. In the example depicted, R may reside on either the 5-membered or the 6-membered ring of the fused ring system. In the formula above, the subscript letter “y” immediately following the R enclosed in parentheses, represents a numeric variable. Unless specified otherwise, this variable can be 0, 1, 2, or any integer greater than 2, only limited by the maximum number of replaceable hydrogen atoms of the ring or ring system.

[0115] For the chemical groups and compound classes, the number of carbon atoms in the group or class is as indicated as follows: “Cn” or “C=n” defines the exact number (n) of carbon atoms in the group / class. “C<n” defines the maximum number (n) of carbon atoms that can be in the group / class, with the minimum number as small as possible for the group / class in question. For example, it is understood that the minimum number of carbon atoms in the groups “alkylc<s”, “cycloalkancdiylc<s”. “heteroarylcss”, and “acylc<s” is one, the minimum number of carbon atoms in the groups “alkcnylc<s”. “alkynylc<g”, and “hctcrocycloalkylc<s” is two, the minimum number of carbon atoms in the group “cycloalky lc<s” is three, and the minimum number of carbon atoms in the groups “arylc<s” and “arcncdiylc<s” is six. “Cn-n'” defines both the minimum (n) and maximum number (n') of carbon atoms in the group. Thus, “alkylc2-io” designates those alkyl groups having from 2 to 10 carbon atoms. These carbon number indicators may precede or follow the chemical groups or class it modifies and it may or may not be enclosed in parenthesis, without signifying any change in meaning. Thus, the terms “C5 olefin”, “C5-olefin”, “olefines)”, and “olefines” are all synonymous. Except as noted below, every carbon atom is counted to determine whether the group or compound falls with the specified number of carbon atoms. For example, the group dihexylamino is an example of a dialkylaminoc=i2 group; however, it is not an example of a dialkylaminoc=6 group. Likewise, phenylethyl is an example of an aralkylc=s group. When any of the chemical groups or compound classes defined herein is modified by the term “substituted”, any carbon atom in the moiety replacing the hydrogen atom is not counted. Thus methoxyhexyl, which has a total of seven carbon atoms, is an example of a substituted alkylci-6. Unless specified otherwise, any chemical group or compound class listed in a claim set without a carbon atom limit has a carbon atom limit of less than or equal to twelve.

[0116] The term “saturated” when used to modify a compound or chemical group means the compound or chemical group has no carbon-carbon double and no carbon-carbon triple bonds, except as noted below. When the term is used to modify an atom, it means that the atom is not part of any double or triple bond. In the case of substituted versions of saturated groups, one or more carbon oxygen double bond or a carbon nitrogen double bond may be present. And when such a bond is present, then carbon-carbon double bonds that may occur as part of ketoenol tautomerism or imine / enamine tautomerism are not precluded. When the term “saturated” is used to modify a solution of a substance, it means that no more of that substance can dissolve in that solution.

[0117] The term “aliphatic” signifies that the compound or chemical group so modified is an acyclic or cyclic, but non-aromatic compound or group. In aliphatic compounds / groups, the carbon atoms can be joined together in straight chains, branched chains, or non-aromatic rings (alicyclic). Aliphatic compounds / groups can be saturated, that is joined by single carboncarbon bonds (alkanes / alkyl), or unsaturated, with one or more carbon-carbon double bonds (alkenes / alkenyl) or with one or more carbon-carbon triple bonds (alkynes / alkynyl). The term “aromatic” signifies that the compound or chemical group so modified has a planar unsaturated ring of atoms with 4n +2 electrons in a fully conjugated cyclic n system. An aromatic compound or chemical group may be depicted as a single resonance structure; however, depiction of one resonance structure is taken to also refer to any other resonance structure. For example:

[0118] Aromatic compounds may also be depicted using a circle to represent the delocalized nature of the electrons in the fully conjugated cyclic n system, two non-limiting examples of which are shown below:

[0119] The term “alkyl” refers to a monovalent saturated aliphatic group with a carbon atom as the point of attachment, a linear or branched acyclic structure, and no atoms other than carbon and hydrogen. The groups -CH3 (Me), -CH2CH3 (Et), -CH2CH2CH3 (n-Pr or propyl), -CH(CH3)2(z'-Pr,!Pr or isopropyl), -CH2CH2CH2CH3 (n-Bu), -CH(CH3)CH2CH3 (sec-butyl), -CH2CH(CH3)2(isobutyl), -C(CH3)3 (tert-butyl, t-butyl, t-Bu or 'Bu). and -CH2C(CH3)3 (neopentyl) are non-limiting examples of alkyl groups. The term “alkanediyl” refers to a divalent saturated aliphatic group, with one or two saturated carbon atom(s) as the point(s) of attachment, a linear or branched acyclic structure, no carbon-carbon double or triple bonds, and no atoms other than carbon and hydrogen. The groups -CH2- (methylene), -CH2CH2-, -CH2C(CH3)2CH2-, and -CH2CH2CH2- are non-limiting examples of alkanediyl groups. The term “alkylidene” refers to the divalent group =CRR' in which R and R' are independently hydrogen or alkyl. Non-limiting examples of alkylidene groups include: =CH2. =CH(CH2CH3), and =C(CH3)2- An “alkane” refers to the class of compounds having the formula H-R, wherein R is alkyl as this term is defined above.

[0120] The term “cycloalkyl” refers to a monovalent saturated aliphatic group with a carbon atom as the point of attachment, said carbon atom forming part of one or more non-aromatic ring structures, no carbon-carbon double or triple bonds, and no atoms other than carbon and hydrogen. Non-limiting examples include: -CH(CH2)2 (cyclopropyl), cyclobutyl, cyclopentyl, or cyclohexyl (Cy). As used herein, the term does not preclude the presence of one or more alkyl groups (carbon number limitation permitting) attached to a carbon atom of the non- aromatic ring structure. The term “cyclo alkanediyl” refers to a divalent saturated aliphatic group with two carbon atoms as points of attachment, no carbon-carbon double or triple bonds, and no atoms other than carbon and hydrogen. The group is a non-limiting example of cycloalkanediyl group. A “cycloalkane” refers to the class of compounds having the formula H-R, wherein R is cycloalkyl as this term is defined above.

[0121] The term “aryl” refers to a monovalent unsaturated aromatic group with an aromatic carbon atom as the point of attachment, said carbon atom forming part of a one or more aromatic ring structures, each with six ring atoms that are all carbon, and wherein the group consists of no atoms other than carbon and hydrogen. If more than one ring is present, the rings may be fused or unfused. Unfused rings are connected with a covalent bond. As used herein, the term aryl does not preclude the presence of one or more alkyl groups (carbon number limitation permitting) attached to the first aromatic ring or any additional aromatic ring present. Non-limiting examples of aryl groups include phenyl (Ph), methylphenyl, (dimethyl)phenyl, -C6H4CH2CH3 (ethylphenyl), naphthyl, and a monovalent group derived from biphenyl (e.g., 4-phenylphenyl). The term “arenediyl” refers to a divalent aromatic group with two aromatic carbon atoms as points of attachment, said carbon atoms forming part of one or more sixmembered aromatic ring structures, each with six ring atoms that are all carbon, and wherein the divalent group consists of no atoms other than carbon and hydrogen. As used herein, the term arenediyl does not preclude the presence of one or more alkyl groups (carbon number limitation permitting) attached to the first aromatic ring or any additional aromatic ring present. If more than one ring is present, the rings may be fused or unfused. Unfused rings are connected with a covalent bond. Non-limiting examples of arenediyl groups include:

[0122] An “arene” refers to the class of compounds having the formula H-R, wherein R is aryl as that term is defined above. Benzene and toluene are non-limiting examples of arenes.

[0123] The term “aralkyl” refers to the monovalent group -alkanediyl-aryl, in which the terms alkanediyl and aryl are each used in a manner consistent with the definitions provided above. Non-limiting examples are: phenylmethyl (benzyl, Bn) and 2-phenyl-ethyl. The term “heterocycloalkyl” refers to a monovalent non-aromatic group with a carbon atom or nitrogen atom as the point of attachment, said carbon atom or nitrogen atom forming part of one or more non-aromatic ring structures, each with three to eight ring atoms, wherein at least one of the ring atoms of the non-aromatic ring structure(s) is nitrogen, oxygen or sulfur, and wherein the heterocycloalkyl group consists of no atoms other than carbon, hydrogen, nitrogen, oxygen and sulfur. If more than one ring is present, the rings are fused. As used herein, the term does not preclude the presence of one or more alkyl groups (carbon number limitation permitting) attached to one or more ring atoms. Also, the term does not preclude the presence of one or more double bonds in the ring or ring system, provided that the resulting group remains non-aromatic. Non-limiting examples of heterocycloalkyl groups include aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydrofuranyl, tetrahydrothiofuranyl, tetrahydropyranyl, pyranyl, oxiranyl, and oxetanyl. The term ‘W-heterocycloalkyl” refers to a heterocycloalkyl group with a nitrogen atom as the point of attachment. A-pyrrolidinyl is an example of such a group.

[0124] The term “acyl” refers to the group -C(O)R, in which R is a hydrogen, alkyl, cycloalkyl, or aryl as those terms are defined above. The groups, -CHO, -C(0)CH3 (acetyl, Ac), -C(O)CH2CH3, -C(O)CH(CH3)2, -C(O)CH(CH2)2, -C(O)C6H5, and -C(O)C6H4CH3are nonlimiting examples of acyl groups. A “thioacyl” is defined in an analogous manner, except that the oxygen atom of the group -C(O)R has been replaced with a sulfur atom, -C(S)R. The term “aldehyde” corresponds to an alkyl group, as defined above, attached to a -CHO group.

[0125] The term “alkoxy” refers to the group -OR, in which R is an alkyl, as that term is defined above. Non-limiting examples include: -OCH3(methoxy), -OCH2CH3(ethoxy), -OCH2CH2CH3, -OCH(CH3)2(isopropoxy), or -OC(CH3)3(tert-butoxy). The terms “cycloalkoxy”, “aryloxy”, “aralkoxy”, “heterocycloalkoxy”, and “acyloxy”, when used without the “substituted” modifier, refers to groups, defined as -OR, in which R is cycloalkyl, aryl, aralkyl, heterocycloalkyl, and acyl, respectively. The term “alkylthio” and “acylthio” refers to the group -SR, in which R is an alkyl and acyl, respectively. The term “alcohol” corresponds to an alkane, as defined above, wherein at least one of the hydrogen atoms has been replaced with a hydroxy group. The term “ether” corresponds to an alkane, as defined above, wherein at least one of the hydrogen atoms has been replaced with an alkoxy group.

[0126] When a chemical group is used with the “substituted” modifier, one or more hydrogen atom has been replaced, independently at each instance, by -OH, -F, -Cl, -Br, -I, -NH2, -N02, -CO2H, -CO2CH3, -CO2CH2CH3, -CN, -SH, -0CH3, -OCH2CH3, -C(O)CH3, -NHCH3, -NHCH2CH3, -N(CH3)2, -C(O)NH2, -C(O)NHCH3, -C(O)N(CH3)2, -OC(O)CH3, -NHC(0)CH3, -S(0)20H, or -S(O)2NH2. For example, the following groups are non-limiting examples of substituted alkyl groups: -CH2OH, -CH2CI, -CF3, -CH2CN, -CH2C(0)0H, -CH2C(O)OCH3, -CH2C(O)NH2, -CH2C(O)CH3, -CH2OCH3, -CH2OC(O)CH3, -CH2NH2, -CH2N(CH3)2, and -CH2CH2CI. The term “haloalkyl” is a subset of substituted alkyl, in which the hydrogen atom replacement is limited to halo (z.e. -F, -Cl, -Br, or -I) such that no other atoms aside from carbon, hydrogen and halogen are present. The group, -CH2CI is a nonlimiting example of a haloalkyl. The term “fluoroalkyl” is a subset of substituted alkyl, in which the hydrogen atom replacement is limited to fluoro such that no other atoms aside from carbon, hydrogen and fluorine are present. The groups -CH2F, -CF3, and -CH2CF3 are nonlimiting examples of fluoroalkyl groups.

[0127] As used herein, the term “functional group” refers to any chemical group or substituent covalently bound to a core structure. Functional groups may include but are not limited to hydrogen, alkyl, cycloalkyl, heterocycloalkyl, aryl, aralkyl, -OH, -F, -Cl, -Br, -I, -NH2, -NO2, -CO2H, -CO2CH3, -CO2CH2CH3, -CN, -SH, -OCH3, -OCH2CH3, -C(O)CH3, -NHCH3, -NHCH2CH3, -N(CH3)2, -C(O)NH2, -C(O)NHCH3, -C(O)N(CH3)2, -OC(O)CH3, -NHC(0)CH3, -S(O)2OH, -S(O)2NH2 or a combination or substituted version of any of these groups.

[0128] The use of the word “a” or “an,” when used in conjunction with the term “comprising” in the claims and / or the specification may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.”

[0129] Throughout this application, the term “about” is used to indicate that a value includes the inherent variation of error for the device, the method being employed to determine the value, or the variation that exists among the study subjects or patients.

[0130] An “active ingredient” (Al) or active pharmaceutical ingredient (API) (also referred to as an active compound, active substance, active agent, pharmaceutical agent, agent, biologically active molecule, or a therapeutic compound) is the ingredient in a pharmaceutical drug that is biologically active.

[0131] The terms “comprise,” “have” and “include” are open-ended linking verbs. Any forms or tenses of one or more of these verbs, such as “comprises,” “comprising,” “has,” “having,” “includes” and “including,” are also open-ended. For example, any method that “comprises,” “has” or “includes” one or more steps is not limited to possessing only those one or more steps and also covers other unlisted steps.

[0132] The term “effective,” as that term is used in the specification and / or claims, means adequate to accomplish a desired, expected, or intended result. “Effective amount,” “Therapeutically effective amount” or “pharmaceutically effective amount” when used in the context of treating a patient or subject with a compound means that amount of the compound which, when administered to a subject or patient for treating or preventing a disease, is an amount sufficient to effect such treatment or prevention of the disease.

[0133] An “excipient” is a pharmaceutically acceptable substance formulated along with the active ingredient(s) of a medication, pharmaceutical composition, formulation, or drug delivery system. Excipients may be used, for example, to stabilize the composition, to bulk up the composition (thus often referred to as “bulking agents,” “fillers,” or “diluents” when used for this purpose), or to confer a therapeutic enhancement on the active ingredient in the final dosage form, such as facilitating drug absorption, reducing viscosity, or enhancing solubility. Excipients include pharmaceutically acceptable versions of antiadherents, binders, coatings, colors, disint egrants, flavors, glidants, lubricants, preservatives, sorbents, sweeteners, and vehicles. The main excipient that serves as a medium for conveying the active ingredient is usually called the vehicle. Excipients may also be used in the manufacturing process, for example, to aid in the handling of the active substance, such as by facilitating powder flowability or non-stick properties, in addition to aiding in vitro stability such as prevention of denaturation or aggregation over the expected shelf life. The suitability of an excipient will typically vary depending on the route of administration, the dosage form, the active ingredient, as well as other factors.

[0134] As used herein, the term “IC50” refers to an inhibitory dose which is 50% of the maximum response obtained. This quantitative measure indicates how much of a particular drug or other substance (inhibitor) is needed to inhibit a given biological, biochemical or chemical process (or component of a process, i.e. an enzyme, cell, cell receptor or microorganism) by half. An “isomer” of a first compound is a separate compound in which each molecule contains the same constituent atoms as the first compound, but where the configuration of those atoms in three dimensions differs.

[0135] As used herein, the term “patient” or “subject” refers to a living mammalian organism, such as a human, monkey, cow, sheep, goat, dog, cat, mouse, rat, guinea pig, or transgenic species thereof. In certain embodiments, the patient or subject is a primate. Non-limiting examples of human patients are adults, juveniles, infants and fetuses.

[0136] As generally used herein “pharmaceutically acceptable” refers to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues, organs, and / or bodily fluids of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit / risk ratio.

[0137] “Pharmaceutically acceptable salts” means salts of compounds disclosed herein which are pharmaceutically acceptable, as defined above, and which possess the desired pharmacological activity. Such salts include acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or with organic acids such as 1 ,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, 2-naphthalenesulfonic acid, 3 -phenylpropionic acid, 4,4'-methylenebis(3-hydroxy-2-ene- 1 -carboxylic acid), 4-methylbicyclo[2.2.2]oct-2-ene-l -carboxylic acid, acetic acid, aliphatic mono- and dicarboxylic acids, aliphatic sulfuric acids, aromatic sulfuric acids, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, carbonic acid, cinnamic acid, citric acid, cyclopentanepropionic acid, ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, heptanoic acid, hexanoic acid, hydroxynaphthoic acid, lactic acid, laurylsulfuric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, muconic acid, o-(4-hydroxybenzoyl)benzoic acid, oxalic acid, p-chlorobenzenesulfonic acid, phenyl-substituted alkanoic acids, propionic acid, p-toluenesulfonic acid, pyruvic acid, salicylic acid, stearic acid, succinic acid, tartaric acid, tertiarybutylacetic acid, trimethylacetic acid, and the like. Pharmaceutically acceptable salts also include base addition salts which may be formed when acidic protons present are capable of reacting with inorganic or organic bases. Acceptable inorganic bases include sodium hydroxide, sodium carbonate, potassium hydroxide, aluminum hydroxide and calcium hydroxide. Acceptable organic bases include ethanolamine, diethanolamine, triethanolamine, tromethamine, A-methylglucamine and the like. It should be recognized that the particular anion or cation forming a part of any salt of this disclosure is not critical, so long as the salt, as a whole, is pharmacologically acceptable. Additional examples of pharmaceutically acceptable salts and their methods of preparation and use are presented in Handbook of Pharmaceutical Salts: Properties, and Use (P. H. Stahl & C. G. Wermuth eds., Verlag Helvetica Chimica Acta, 2002).

[0138] A “pharmaceutically acceptable carrier,” “drug carrier,” or simply “carrier” is a pharmaceutically acceptable substance formulated along with the active ingredient medication that is involved in carrying, delivering and / or transporting a chemical agent. Drug carriers may be used to improve the delivery and the effectiveness of drugs, including for example, controlled-release technology to modulate drug bioavailability, decrease drug metabolism, and / or reduce drug toxicity. Some drug carriers may increase the effectiveness of drug delivery to the specific target sites. Examples of carriers include: liposomes, microspheres (e.g., made of poly(lactic-co-glycolic) acid), albumin microspheres, synthetic polymers, nanofibers, protein-DNA complexes, protein conjugates, erythrocytes, virosomes, and dendrimers.

[0139] A “pharmaceutical drug” (also referred to as a pharmaceutical, pharmaceutical preparation, pharmaceutical composition, pharmaceutical formulation, pharmaceutical product, medicinal product, medicine, medication, medicament, or simply a drug, agent, or preparation) is a composition used to diagnose, cure, treat, or prevent disease, which comprises an active pharmaceutical ingredient (API) (defined above) and optionally contains one or more inactive ingredients, which are also referred to as excipients (defined above).

[0140] “Prevention” or “preventing” includes: (1) inhibiting the onset of a disease in a subject or patient which may be at risk and / or predisposed to the disease but does not yet experience or display any or all of the pathology or symptomatology of the disease, and / or (2) slowing the onset of the pathology or symptomatology of a disease in a subject or patient which may be at risk and / or predisposed to the disease but does not yet experience or display any or all of the pathology or symptomatology of the disease.

[0141] A “stereoisomer” or “optical isomer” is an isomer of a given compound in which the same atoms are bonded to the same other atoms, but where the configuration of those atoms in three dimensions differs. “Enantiomers” are stereoisomers of a given compound that are mirror images of each other, like left and right hands. “Diastereomers” are stereoisomers of a given compound that are not enantiomers. Chiral molecules contain a chiral center, also referred to as a stereocenter or stereogenic center, which is any point, though not necessarily an atom, in a molecule bearing groups such that an interchanging of any two groups leads to a stereoisomer. In organic compounds, the chiral center is typically a carbon, phosphorus or sulfur atom, though it is also possible for other atoms to be stereocenters in organic and inorganic compounds. A molecule can have multiple stereocenters, giving it many stereoisomers. In compounds whose stereoisomerism is due to tetrahedral stereogenic centers (e.g., tetrahedral carbon), the total number of hypothetically possible stereoisomers will not exceed 2n, where n is the number of tetrahedral stereocenters. Molecules with symmetry frequently have fewer than the maximum possible number of stereoisomers. A 50:50 mixture of enantiomers is referred to as a racemic mixture. Alternatively, a mixture of enantiomers can be enantiomerically enriched so that one enantiomer is present in an amount greater than 50%. Typically, enantiomers and / or diastereomers can be resolved or separated using techniques known in the art. It is contemplated that that for any stereocenter or axis of chirality for which stereochemistry has not been defined, that stereocenter or axis of chirality can be present in its R form, S form, or as a mixture of the R and S forms, including racemic and non-racemic mixtures. As used herein, the phrase “substantially free from other stereoisomers” means that the composition contains < 15%, more preferably < 10%, even more preferably < 5%, or most preferably < 1% of another stereoisomer(s).

[0142] “Treatment” or “treating” includes (1) inhibiting a disease in a subject or patient experiencing or displaying the pathology or symptomatology of the disease (e.g., arresting further development of the pathology and / or symptomatology), (2) ameliorating a disease in a subject or patient that is experiencing or displaying the pathology or symptomatology of the disease (e.g., reversing the pathology and / or symptomatology), and / or (3) effecting any measurable decrease in a disease or symptom thereof in a subject or patient that is experiencing or displaying the pathology or symptomatology of the disease.

[0143] The term “ubiquitin ligase ligand” or “E3 ligase ligand” refers to a chemical group capable of binding ubiquitin ligase. Ubiquitin ligase (also called an E3 ubiquitin ligase) is a protein that recruits an E2 ubiquitin-conjugating enzyme that has been loaded with ubiquitin, recognizes a protein substrate, and assists or directly catalyzes the transfer of ubiquitin from the E2 to the protein substrate. The ubiquitin is attached to a lysine on the target protein by an isopeptide bond. E3 ligases interact with both the target protein and the E2 enzyme, and so impart substrate specificity to the E2. Commonly, E3s polyubiquitinate their substrate with Lys48-linked chains of ubiquitin, targeting the substrate for destruction by the proteasome. However, one of skill in the art recognizes that many other types of linkages are possible and that each may alter a protein's activity, interactions, or localization. Ubiquitination by E3 ligases regulates diverse areas such as cell trafficking, DNA repair, and signaling and is of profound importance in cell biology. E3 ligases are also key players in cell cycle control, mediating the degradation of cyclins, as well as cyclin dependent kinase inhibitor proteins. The human genome encodes over 600 putative E3 ligases, allowing for tremendous diversity in substrates. Non-limiting examples of ubiquitin ligase ligands include the von Hippel-Lindau (VHL) ligand, a cIAPl ligand, a MDM2 ligand, a CRBN ligand, a CUL2 ligand, or other ligand which binds to one or more of the proteins of the ubiquitin protein complex especially the E3 component of this complex.

[0144] Ubiquitin ligase (also called an E3 ubiquitin ligase or E3 ligase) is a protein that recruits an E2 ubiquitin-conjugating enzyme that has been loaded with ubiquitin, recognizes a protein substrate, and assists or directly catalyzes the transfer of ubiquitin from the E2 to the protein substrate. The ubiquitin is attached to a lysine on the target protein by an isopeptide bond. E3 ligases interact with both the target protein and the E2 enzyme, and so impart substrate specificity to the E2. Commonly, E3s polyubiquitinate their substrate with Lys48-linked chains of ubiquitin, targeting the substrate for destruction by the proteasome. However, one of skill in the art recognizes that many other types of linkages are possible and that each may alter a protein's activity, interactions, or localization. Ubiquitination by E3 ligases regulates diverse areas such as cell trafficking, DNA repair, and signaling and is of profound importance in cell biology. E3 ligases are also key players in cell cycle control, mediating the degradation of cyclins, as well as cyclin dependent kinase inhibitor proteins. The human genome encodes over 600 putative E3 ligases, allowing for tremendous diversity in substrates. Non-limiting examples of ubiquitin ligase ligands include the von Hippel-Lindau (VHL) ligand, a cIAPl ligand, a MDM2 ligand, a CRBN ligand, a CUL2 ligand, or other ligand which binds to one or more of the proteins of the ubiquitin protein complex especially the E3 component of this complex.

[0145] The term “unit dose” refers to a formulation of the compound or composition such that the formulation is prepared in a manner sufficient to provide a single therapeutically effective dose of the active ingredient to a patient in a single administration. Such unit dose formulations that may be used include but are not limited to a single tablet, capsule, or other oral formulations, or a single vial with a syringeable liquid or other injectable formulations.

[0146] The above definitions supersede any conflicting definition in any reference that is incorporated by reference herein. The fact that certain terms are defined, however, should not be considered as indicative that any term that is undefined is indefinite. Rather, all terms used are believed to describe the disclosure in terms such that one of ordinary skill can appreciate the scope and practice the present disclosure.

[0147] I. Examples

[0148] The following examples are included to demonstrate preferred embodiments of the disclosure. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the disclosure, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the disclosure.

[0149] Example 1 - Synthetic Methodology and Characterization Data

[0150] A. Synthesis of MDM2 PROTACs

[0151] General Procedure for Synthesis of MDM2-VHL PROTACs:

[0152] Step-1 : A mixture of VHL-amine I (0.21 mmol, 1.0 equiv), acid linker- 1 II (0.23 mmol, 1.1 equiv), HATU (0.25 mmol, 1.2 equiv) and z-P NEt (0.63 mmol, 3.0 equiv) in DCM (3.0 mL) was stirred at room temperature for 1 h. The mixture was then diluted with DCM and organic layer was separated. The combined organic layers were washed with saturated aqueous NaHCCL solution, dried over Na2SC>4, and concentrated under reduced pressure to give a residue, which was purified by silica gel flash column chromatography with DCM: MeOH (9: 1 - 4:1) to afford the compound III.

[0153] Step-2: 4N HC1 in dioxane (1.2 mL) was added dropwise to a mixture of III (~ 0.18 mmol) in DCM (3.6 mL) and the resulting suspension was allowed to stir at room temperature for 1 h. The mixture was then concentrated, and the obtained solid residue was washed with diethyl ether and dried under high vacuum to afford product IV as HC1 salt, which was used in the next step without further purification.

[0154] Step-3: To a room temperature solution of IV (~ 0.18 mmol, 1.0 equiv), acid linker-2 V (0.18 mmol, 1.0 equiv) and z'-PnNEt (0.54 mmol, 3.0 equiv) in DCM (2.0 mL), HATU (0.22 mmol, 1.2 equiv) was added. The resulting solution was stirred for 1 h. The reaction mixture was then diluted with DCM and the organic phase was separated followed by washed with a saturated aqueous NaHCCL solution. After drying the organic phase over sodium sulfate and evaporation of the solvent under reduced pressure, the obtained residue was purified by silica gel flash column chromatography with DCM: MeOH (9:1 - 4:1) to get the compound VI.

[0155] Step-4: Compound VI (~ 0.14 mmol) was dissolved in DCM (2.7 mL) and 4N HC1 in dioxane (900 pL) was added dropwise at room temperature and stirring was continued for 1 h. After the completion of the reaction, the solvent was removed under reduced pressure, and the residue obtained was washed with diethyl ether and dried under vacuum to afford product VII as HC1 salt, which was used in the next step without further purification.

[0156] Step-5: To a stirred solution MDM2 acid VIII (0.14 mmol, 1.0 equiv) in DCM (2.5 mL) was added HATU (0.17 mmol, 1.2 equiv), z'-P NEt (0.42 mmol, 3.0 equiv) and the corresponding amine salt VII (~ 0.14 mmol). The reaction mixture was stirred at room temperature for 1 h. After completion of reaction (as indicated by LCMS), organic phase was extracted in DCM and the combined organic phase was washed with a saturated aqueous solution of NaHCCL and dried over Na2SC>4. After evaporation of the solvent in vacuum, the residue was purified by silica gel flash column chromatography with DCM: MeOH (9:1 - 7:3) to yield the desired compound.

[0157] rac-l-(2-(4-((4S,5R)-2-(4-tert-butyl-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)-4,5- dimethyl-4,5-dihydro- 1 / / -imidazole- 1 -carbonyl )piperazin-l-yl )acetyl ((2S,4R)-4-hyd-roxy-2-(4-(4-methylthiazol-5-yl)benzylcarbamoyl)pyrrolidin-l-yl)-3,3- dimethyl-l-oxobu-tan-2-ylamino)-2-oxo-l-phenylethyl)piperidine-4-carboxamide (MKM-01-031): Compound MKM-01-031 was synthesized using general procedure (steps 1- 5) and racemic MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 96.1%. LCMS: m / z calculated for C72H86CI2N10O8S = 1320.57; found [M+2H / 2]+= 661.49. rac-l-(2-(4-((4S,5R)-2-(4-tert-butyl-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)-4,5- dimethyl-4,5-dihydro- 1 / / -i mid azole- 1 -carbonyl )piperazin-l-yl )acetyl)-V-( 1-((S)-1- ((2S,4R)-4-hydr-oxy-2-(4-(4-methylthiazol-5-yl)benzylcarbamoyl)pyrrolidin-l-yl)-3,3- dimethyl-l-oxobutan-2-yl-carbamoyl)cyclopropyl)piperidine-4-carboxamide (MKM-01- 038): Compound MKM-01-038 was synthesized using general procedure (steps 1-5) and racemic MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 97.3%. LCMS: m / z calculated for C68H84Cl2N10O8S = 1270.56; found [M+2H / 2]+= 636.40.

[0158] rac-l-(2-(4-((4S,5R)-2-(4-(tert-butyl)-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)- 4, 5-dimethyl-4,5-dihydro- 1 / / -imidazole- l-carbonyl)piperazin- 1-yl )acetyl )-N-( 1-(((S)- 1- ((2S,4R)-4-hydr-oxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-l-yl)-3,3- dimethyl- 1-oxobutan -2-yl)-carbamoyl)cyclopentyl)piperidine-4-carboxamide (MKM- 01-042): Compound MKM-01-042 was synthesized using general procedure (steps 1-5) and racemic MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 95.2%. LCMS: m / z calculated for C68H84Cl2N10O8S = 1298.59; found [M+2H / 2]+= 650.49. rac-l-(2-(4-((4S,5R)-2-(4-tert-butyl-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)-4,5- dimethyl-4,5-dihydro- 1 / / -i mid azole- 1 -carbonyl )piperazin-l-yl)acetyl)-V-((.S)-l -((2S,4R)- 4-hydroxy-2-(4-(4-methylthiazol-5-yl)benzylcarbamoyl)pyrrolidin-l-yl)-3,3-dimethyl-l- oxobutan-2-yl)pipe-ridine-4-carboxamide (MKM-01-043): Compound MKM-01-043 was synthesized using general procedure (steps 1, 2 & 5) and racemic MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 98%. LCMS: m / z calculated for C64H79CI2N9O7S = 1187.52; found [M+H]+= 1188.73.

[0159] rac-l-(2-(4-((4S,5R)-2-(4-tert-butyl-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)-4,5- dimethyl-4,5-dihydro- 1 / / -i mid azole- 1 -carbonyl )piperazin-l-yl )acetyl )-.V-((.S')-l-((.S')-l- ((2S,4R)-4-hydroxy-2-(4-(4-methylthiazol-5-yl)benzylcarbamoyl)pyrrolidin-l-yl)-3,3- dimethyl-l-oxo-butan-2-ylamino)-l-oxo-3-phenylpropan-2-yl)piperidine-4-carboxamide (MKM-01-077): Compound MKM-01-077 was synthesized using general procedure (steps 1- 5) and racemic MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 97.6%. LCMS: m / z calculated for CvsHggCLNioOgS = 1334.59; found [M+2H / 2]+= 668.79. rac-(2S,4R)-l-((S)-2-(l-(2-(2-(2-(4-((4S,5R)-2-(4-te / 7-butyl-2-ethoxyphenyl)-4,5- bis(4-chloro-phenyl)-4,5-dimethyl-4,5-dihydro-177-imidazole-l-carbonyl)piperazin-l- yl)acetamido)-ethoxy)acetamido)cyclopropanecarboxamido)-3,3-dimethylbutanoyl)-4- hydroxy-Ar-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (MKM-01-084):

[0160] Compound MKM-01-084 was synthesized using general procedure (steps 1-5) and racemic MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 95.7%. LCMS: m / z calculated for C66H82Q2N10O9S = 1260.54; found [M+2H / 2]+= 631.40. Chiral (2S,4fl)-l-((S)-2-( l-(2-(2-(2-(4-((4S,5R)-2-(4-(tert-butyl)-2-ethoxyphenyl)-

[0161] 4,5-bis(4-chloro-phenyl)-4,5-dimethyl-4,5-dihydro-l / / -imidazole-l-carbonyl)piperazin- l-yl)acetamido)-ethoxy)acetamido)cyclopropane-l-carboxamido)-3,3- dimethylbutanoyl)-4-hydroxy-Ar-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2- carboxamide (MKM-01-099): Compound MKM-01-099 was synthesized using general procedure (steps 1-5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 94.9%. LCMS: m / z calculated for C66H82CI2N10O9S = 1260.54; found [M+2H / 2]+= 631.30.

[0162] Chiral l-(2-(4-((4S,5R)-2-(4-(tert-butyl)-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)-

[0163] 4,5-dime-thyl-4,5-dihydro- 1 / / -imidazole- 1 -carbonyl )piperazin- 1-yl )acetyl)-V-( 1-(((S)-1- ((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-l-yl)-3,3- dimethyl-l-oxo-butan-2-yl)-carbamoyl)cyclopropyl)piperidine-4-carboxamide (MKM- 01-103): Compound MKM-01-103 was synthesized using general procedure (steps 1-5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 96.8%. LCMS: m / z calculated for C68H84Cl2N10O8S = 1270.56; found [M+2H / 2]+= 636.40.

[0164] Chiral l-(2-(4-((4S,5R)-2-(4-(tert-butvl)-2-ethoxyphenvl)-4,5-bis(4-chlorophenvl)-

[0165] 4, 5-dime-thvl-4,5-dihvdro- 1 / / -imidazole- 1 -carbonyl )piperazin-l-yl)acetyl)-V-((.S)-l- (((.S)-l-((2.S,4R)-4-hvdroxy-2-((4-(4-inethvlthiazol-5-yl)beiizyl)carbamoyl)pyrrolidin- 1- yl)-3,3-dimethyl-l-oxobutan-2-yl)amino)-l-oxo-3-phenylpropan-2-yl)piperidine-4- carboxamide (MKM-01-106): Compound MKM-01-106 was synthesized using general procedure (steps 1-5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 97.8%. LCMS: m / z calculated for C68H84Cl2N10O8S = 1334.59; found [M+2H / 2]+= 668.49.

[0166] Chiral l-(2-(4-((4S,5R)-2-(4-(tert-butyl)-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)- 4, 5-dime-thyl-4,5-dihydro- l / / -imidazole- 1 -carbonyl )piperazin- 1-yl (acetyl )- V-(( / ?(- 1- (((S)-l-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-l- yl)-3,3-dimethyl-l-oxobutan-2-yl)-amino)-l-oxo-3-phenylpropan-2-yl)piperidine-4- carboxamide (MKM-01-125): Compound MKM-01-125 was synthesized using general procedure (steps 1-5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 93.4%. LCMS: m / z calculated for CvsHssCLNioOgS = 1334.59; found [M+2H / 2]+= 668.51.

[0167] Chiral l-(2-(4-((4S,5R)-2-(4-(tert-butyl)-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)- 4,5-dime-thyl-4,5-dihydro- 1 / / -imidazole- 1 -carbonyl )piperazin-l-yl (acetyl )-V-( 1-(((S)-1- ((2.S,4R)-4-hydroxy-2-((4-(4-niethylthiazol-5-yl)benzyl (carbamoyl (pyrrolidin- l-yl)-3, 3- dimethyl-l-oxo-butan-2-yl)carbamoyl)cyclopropyl)piperidine-3-carboxamide (MKM- 01-127): Compound MKM-01-127 was synthesized using general procedure (steps 1-5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 99.1%. LCMS: m / z calculated for C68H84Cl =2N 11207O08.S56; found [M+2H / 2]+=

[0168] 636.52.

[0169] Chiral l-(2-(4-((4S,5R)-2-(4-(tert-butyl)-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)- 4, 5-dime-thvl-4,5-dihvdro- 1 / / -imidazole- 1 -carbonyl )piperazin-l-yl)acetyl)-V-((.S)-l-

[0170] ((2S,4R)-4-hydr-oxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-l-yl)-3,3- dimethyl-l-oxobu-tan-2-yl)piperidine-4-carboxamide (MKM-01-130): Compound MKM- 01-130 was synthesized using general procedure (steps 1, 2 & 5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 98.8%. LCMS: m / z calculated for C64H79CI2N9O7S = 1187.52; found [M+H]+= 1188.72.

[0171] Chiral l-(2-(4-((4S,5R)-2-(4-(tert-butyl)-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)- 4, 5-dime-thvl-4,5-dihvdro- 1 / / -imidazole- 1 -carbonyl )piperazin-l-yl)acetyl)-V-((.S)-l- ((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-l-yl)-3,3- dimethyl-l-oxobutan-2-yl)piperidine-3-carboxamide (MKM-01-137): Compound MKM- 01-137 was synthesized using general procedure (steps 1, 2 & 5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 99.2%. LCMS: m / z calculated for C64H79CI2N9O7S = 1187.52; found [M+H]+= 1188.72.

[0172]

[0173] Chiral (2.S,4R)-l-((.S)-2-(2-( l-(2-(4-((4S,5R)-2-(4-(tert-butyl)-2-ethoxyphenyl)-4,5- bis(4-chlo-ro-phenvl)-4,5-dimethvl-4,5-dihvdro- l / / -imidazole- 1 -carbonyl )piperazin- 1- yl)acetyl) piperidin-4-yl)acetamido)-3,3-dimethylbutanoyl)-4-hydroxyW-(4-(4- methylthiazol-5-yl) benzyl)pyro-lidine-2-carboxamide (MKM-01-140): Compound MKM-01-140 was synthesized using general procedure (steps 1, 2 & 5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 97.4%. LCMS: m / z calculated for C65H81CI2N9O7S = 1201.54; found [M+2H / 2]+= 601.93. Chiral l-(2-(4-((4S,5R)-2-(4-(tert-butvl)-2-ethoxyphenvl)-4,5-bis(4-chlorophenvl)-

[0174] 4, 5-dime-thvl-4,5-dihvdro- 1 / / -imidazole- 1 -carbonyl )piperazin-l-yl)acetyl)-V-((.S)-l- ((2S,4R)-4-hydr-oxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-l-yl)-3,3- dimethyl-l-oxobutan -2-yl)azepane-4-carboxamide (MKM-01-143): Compound MKM-

[0175] 01-143 was synthesized using general procedure (steps 1, 2 & 5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 99.2%. LCMS: m / z calculated for C65H81CI2N9O7S = 1201.54; found [M+2H / 2]+= 601.93. Chiral (2.S,4R)-l-((.S)-2-(2-(2-(2-(4-((4S,5R)-2-(4-(tert-butyl)-3-ethoxyphenyl)-4,5- bis(4-chlo-rophenyl)-4,5-dimethyl-4,5-dihydro-177-imidazole-l-carbonyl)piperazin-l- yl)acetamido) ethoxy)acetamido)-3,3-dimethylbutanoyl)-4-hydroxyW-(4-(4- methylthiazol-5-yl)benzyl) pyrrolidine-2-carboxamide (MKM-01-149): Compound MKM-01-149 was synthesized using general procedure (steps 1, 2 & 5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 98.6%. LCMS: m / z calculated for C62H77CI2N9O8S = 1177.50; found [M+H]+= 1178.72.

[0176] Chiral (2.S,4R)-l-((.S)-2-(3-( l-(2-(4-((4S,5R)-2-(4-(tert-butyl)-2-ethoxyphenyl)-4,5- bis(4-chlo-rophe-nyl)-4,5-dimethyl-4,5-dihydro- l / / -imidazole- 1 -carbonyl )piperazin- 1- yl)acetyl) piperidiii-4-yl)propaiianiido)-3,3-diniethvlbutanoyl)-4-hvdroxy-V-(4-(4- methylthiazol-5-yl)benzyl)-pyrrolidine-2-carboxamide (MKM-01-153): Compound MKM-01-153 was synthesized using general procedure (steps 1, 2 & 5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 98.9%. LCMS: m / z calculated for C66H83C12N9O7S = 1215.55; found [M+2H / 2]+= 608.83.

[0177] Chiral 2-(2-(4-((4S,5R)-2-(4-(tert-butyl)-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)-

[0178] 4, 5-dime-thvl-4,5-dihvdro- 1 / / -imidazole- 1 -carbonyl )piperazin-l-yl)acetyl)-V-((.S)-l-

[0179] ((2S,4R)-4-hydr-oxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-l-yl)-3,3- dimethyl-l-oxobutan-2-yl)-2-azaspiro[3.3]heptane-6-carboxamide (MKM-01-159):

[0180] Compound MKM-01-159 was synthesized using general procedure (steps 1, 2 & 5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 97%. LCMS: m / z calculated for C65H79CI2N9O7S = 1199.52; found [M+2H / 2]+=

[0181] 600.93.

[0182] Chiral l-(2-(4-((4S,5R)-2-(4-(tert-butyl)-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)- 4, 5-diine-thvl-4,5-dihvdro- 1 / / -imidazole- 1 -carbonyl )pipeiazin-l-yl)acetyl)-V-((.S)-l- ((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-l-yl)-3,3- dimethyl-l-oxobutan-2-yl)-4-methylpiperidine-4-carboxamide (MKM-01-169):

[0183] Compound MKM-01-169 was synthesized using general procedure (steps 1, 2 & 5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 97.7%. LCMS: m / z calculated for C65H81CI2N9O7S = 1201.54; found [M+2H / 2]+= 602.10.

[0184] Chiral (2.S,4R)-l-((.S)-2-(3-( l-(2-(4-((4S,5R)-2-(4-(tert-butyl)-2-ethoxyphenyl)-4,5- bis(4-chl-oro-phenyl)-4,5-dimethyl-4,5-dihydro- l / / -imidazole- 1 -carbonyl )piperazin- 1- yl)acetyl) piperidiii-4-yl)propaiianiido)-3,3-diniethvlbutaiiovl)-4-hvdroxy- V-((.S)-l-(4-(4- methyl-thiazol-5-yl)-phenyl)ethyl)pyrrolidine-2-carboxamide (MKM-01-217):

[0185] Compound MKM-01-217 was synthesized using general procedure (steps 1, 2 & 5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 99.9%. LCMS: m / z calculated for C67H85C12N9O7S = 1229.57; found [M+2H / 2]+= 615.90.

[0186] Chiral l-(2-(4-((4S,5R)-2-(4-(tert-butyl)-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)- 4, 5-diine-thvl-4,5-dihvdro- 1 / / -imidazole- 1 -carbonyl )piperazin-l-yl)acetyl)-N-((S)-l- ((2S,4R)-4-hydr-oxy-2-(((S)-l-(4-(4-methylthiazol-5- yl)phenyl)ethyl)carbamoyl)pyrrolidin-l-yl)-3,3-dime-thyl-l-oxobutan-2-yl)azepane-4- carboxamide (MKM-01-219): Compound MKM-01-219 was synthesized using general procedure (steps 1 , 2 & 5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 99.9%. LCMS: m / z calculated for C66H83Cl2N9O7S = 1215.55; found [M+2H / 2]+= 609.10.

[0187] Chiral 2-(2-(4-((4S,5R)-2-(4-(tert-butvl)-2-ethoxvphenvl)-4,5-bis(4-chlorophenvl)-

[0188] 4,5-dime-thyl-4,5-dihydro-177-imidazole-l-carbonyl)piperazin-l-yl)acetyl)W-((S)-l- ((2S,4R)-4-hydr-oxy-2-(((S)-l-(4-(4-methylthiazol-5- yl)phenyl)ethyl)carbamoyl)pyrrolidin-l-yl)-3,3-dime-thyl-l-oxobutan-2-yl)-2- azaspiro[3.3]heptane-6-carboxamide (MKM-01-227): Compound MKM-01-227 was synthesized using general procedure (steps 1, 2 & 5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 99.9%. LCMS: m / z calculated for C66H8IC12N9O7S = 1213.54; found [M+2H / 2]+= 608.00.

[0189]

[0190] Chiral l-(2-(4-((4S,5R)-2-(4-( r / -butyl)-2-ethoxyphenyl)-4,5-bis(4-chlorophenyl)- 4, 5-diine-thvl-4,5-dihvdro- 1 / / -imidazole- 1 -carbonyl )piperazin-l-yl)acetyl)-V-((.S)-l- ((2S,4R)-4-hydroxy-2-(((S)-l-(4-(4-methylthiazol-5- yl)phenyl)ethyl)carbamoyl)pyrrolidin-l-yl)-3,3-dimethyl-l-oxobutan-2-yl)piperidine-3- carboxamide (MKM-01-271): Compound MKM-01-271 was synthesized using general procedure (steps 1 , 2 & 5) and an active enantiomer of MDM2 ligand (RG7112 acid analogue) was used in step-5. Purity by LCMS: 99.6%. LCMS: m / z calculated for C65H81Q2N9O7S = 1201.54; found [M+2H / 2]+= 602.10.

[0191] EXAMPLE 2 - ANALYSIS AND ACTIVITY OF MDM2 DEGRADER

[0192] A. Mdm2 Western Blot Quantification and Protocol

[0193] The Mdm2 protein level, determined by Western Blot, was quantified after compound treatment for 16-hours in NanoLuc-MDM2 expressing 293T cells. The compounds were diluted using a 5-fold dilution scheme starting from 1000 nM, evaluating Mdm2 protein levels after a 16-hour treatment with compound concentrations of 1000 nM, 200 nM, 40 nM, 8 nM, and 0 nM (untreated DMSO control). The graph was generated using GraphPad PRISM 10. This data is shown in Fig. 1.

[0194] Some of the degraders were tested using standard Western Blotting, gel electrophoresis, to determine the extent to which the compounds lead to decreased Mdm2 protein expression. For Immunoblotting, cells were lysed with lx cell lysis buffer (Cell Signaling Technology, Cat#9803) containing protease inhibitor and phosphatase inhibitor (Roche). The lysates were incubated on ice for 10 minutes followed by removal of the lysed cells and sonication for 10 seconds, and then centrifugation at 13,000g in a benchtop centrifuge at 4°C for 15 minutes. The protein concentration was determined in the samples using the Pierce BCA Protein Kit (Thermo Scientific Cat#23227). The solid protein was denatured at 95°C for 5 minutes in NuPAGE LDS Sample Buffer (4X) (Thermo Scientific Inc) containing 50 mM DTT. Samples were loaded (30 pg) to 4-12% NuPAGE Bis-Tris Gels (Thermo Scientific) and run at 60V for 30 minutes, then 100 V for 2 hours, with lx NuPAGE MOPS SDS Running Buffer (20X Stock). Following electrophoresis, the samples were transferred onto polyvinylidene fluoride (PVDF) membranes via an iBlot-2 system (Thermo Fisher Scientific Inc; 20V for 10 minutes) and blocked using Intercept Blocking Buffer (IBB) (Ei-COR) for 1-2 hours on a rocker. The primary antibodies were treated in IBB containing 0.2% tween- 20 overnight at 4°C on the rocker. The following primary antibodies used at 1:1000 dilution were purchased from Cell Signaling Technology: Mdm2 (Catalog number CST#86934, rabbit, 60 kDa) and GAPDH (Catalog number CST#97166, mouse, 36 kDa). The secondary antibodies, IRDye 800CW, and IRDye 680RD, were used at 1:5000 dilution, were purchased from Li-Cor. The Western blot images were recorded using the Li-Cor Odyssey M blot imager and the Western blot quantification was done using Empiria Studio software (version 3.0). These data are shown in FIGS. 2-4.

[0195] REFERENCES

[0196] The following references, to the extent that they provide exemplary procedural or other details supplementary to those set forth herein, are specifically incorporated herein by reference:

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Claims

WHAT IS CLAIMED:

1. A compound of the formula:wherein:Ra, Rb, and Rcare each independently hydrogen, alkylccn, or substituted alkylc<12;Ri, R2, R2', R3, and R3' are each independently alkylc is, substituted alkylc is, arylccis, or substituted arylc is;Y is ((CH2)xO)y, wherein x is 1, 2, or 3 and y is 1, 2, or 3 or Y is taken together with Rato form a heterocycloalkaneccn or substituted hctcrocyc loal k ancc i 2 ;A is a covalent bond, C(O), alkanediylccs, or substituted alkancdiylc s; n is 1, 2, or 3;R4 is alkylccs, substituted alkylccs, arylccs, substituted arylccs, aralkylccs, substituted aralkylccs, or taken together with R4 to form a cycloalkaneccs or substituted cycloalkaneccs group;R4' is hydrogen or taken together with R4 to form a cycloalkaneccs or substituted cycloalkaneccs group;Rs is hydroxy, alkoxycce, or substituted alkoxycce; andRe and R7 are each independently selected from alkylccs or substituted alkylccs; or a pharmaceutically acceptable salt thereof.The compound of claim 1 further defined as:wherein:Rb and Rcare each independently hydrogen, alkylc<i2, or substituted alkylc<i2;Ri, R2, R2', R3, and R3' are each independently alkylc<i2, substituted alkylc<i2, arylc<i8, or substituted arylc<is; m is 0, 1, 2, or 3;A is a covalent bond, C(O), alkanediylc<8, or substituted alkanediylc<s; n is 1, 2, or 3;R4 is alkylc<8, substituted alkylc<8, arylc<8, substituted arylc<8, aralkylc<8, substituted aralkylc<8, or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group;R4' is hydrogen or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group;R5 is hydroxy, alkoxyc<6, or substituted alkoxyc<e; andRe and R7 are each independently selected from alkylc<8 or substituted alkylc<s; or a pharmaceutically acceptable salt thereof.The compound of either claim 1 or claim 2 further defined as:wherein:Rb and Rcare each independently hydrogen, alkylc<i2, or substituted alkylc<i2; m is 0, 1, 2, or 3;A is a covalent bond, C(O), alkanediylc<8, or substituted alkancdiylc s; n is 1, 2, or 3;R4 is alkylc<8, substituted alkylc<8, arylc<8, substituted arylc<8, aralkylc<8, substituted aralkylc<8, or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group;R4' is hydrogen or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group;Rs is hydroxy, alkoxyc<6, or substituted alkoxyc<e; andRe and R7 are each independently selected from alkylc<8 or substituted alkylc<s; or a pharmaceutically acceptable salt thereof.The compound according to any one of claims 1-3 further defined as:wherein:Rb and Rcare each independently hydrogen, alkylc<i2, or substituted alkylc<i2; m is 0, 1, 2, or 3;A is a covalent bond, C(O), alkanediylc<8, or substituted alkancdiylc s; n is 1, 2, or 3;R4 is alkylc<8, substituted alkylc<8, arylc<8, substituted arylc<8, aralkylc<8, substituted aralkylc<8, or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group;R4' is hydrogen or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group;Rs is hydroxy, alkoxyc<6, or substituted alkoxyc<e; andRe and R7 are each independently selected from alkylc<8 or substituted alkylc<s; or a pharmaceutically acceptable salt thereof.

5. The compound according to any one of claims 1-4 further defined as:wherein:Rb and Rcare each independently hydrogen, alkylc<i2, or substituted alkylc<i2; m is 0, 1, 2, or 3;A is a covalent bond, C(O), alkanediylc<8, or substituted alkancdiylc s; n is 1, 2, or 3;R4 is alkylc<8, substituted alkylc<8, arylc<8, substituted arylc<8, aralkylc<8, substituted aralkylc<8, or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group;R4' is hydrogen or taken together with R4 to form a cycloalkanec<8 or substituted cycloalkanec<8 group; andRe is alkylc<8 or substituted alkylc<s; or a pharmaceutically acceptable salt thereof.

6. The compound of claim 1, wherein Rais hydrogen.

7. The compound of either claim 1 or claim 6, wherein Y is ((CH2)xO)y, wherein x is 1,2, or 3 and y is 1, 2, or 3.

8. The compound of claim 7, wherein x is 1.

9. The compound of claim 7, wherein x is 2.

10. The compound according to any one of claims 7-9, wherein y is 1.

11. The compound of claim 1 , wherein Y is taken together with Rato form a heterocycloalkanec<i2 or substituted heterocycloalkanec<i2.

12. The compound of claim 11, wherein the heterocycloalkanec<i2 is a pyrrolidine, a piperidine, an azepane, an azocane or an alkyl substituted version of any of these compounds.

13. The compound according to any one of claims 1, 2, and 6-12, wherein Ri is substituted arylc<i8-14. The compound of claim 13, wherein Ri is 2-ethoxy-4-t-butylphenyl.

15. The compound according to any one of claims 1, 2, and 6-14, wherein R2 and R2' are different.

16. The compound according to any one of claims 1, 2, and 6-15, wherein the carbon atom to which R2 and R2' are attached is in the R configuration.

17. The compound according to any one of claims 1, 2, and 6-15, wherein the carbon atom to which R2 and R2' are attached is in the S configuration.

18. The compound according to any one of claims 1, 2, and 6-17, wherein R2 is substituted arylc<is.

19. The compound of claim 18, wherein R2 is 4-chlorophenyl.

20. The compound according to any one of claims 1, 2, and 6-19, wherein R2' is alkylc<8-21. The compound of claim 20, wherein R2' is methyl.

22. The compound according to any one of claims 1, 2, and 6-21, wherein R3 and R3' are different.

23. The compound according to any one of claims 1, 2, and 6-22, wherein the carbon atom to which R3 and R3' are attached is in the R configuration.

24. The compound according to any one of claims 1, 2, and 6-22, wherein the carbon atom to which R3 and R3' are attached is in the S configuration.

25. The compound according to any one of claims 1, 2, and 6-24, wherein R3 is substituted arylc<i8-26. The compound of claim 25, wherein R3 is 4-chlorophenyl.

27. The compound according to any one of claims 1, 2, and 6-26, wherein R3' is alkylc<8-28. The compound of claim 27, wherein R3' is methyl.

29. The compound according to any one of claims 1-28, wherein A is a covalent bond.

30. The compound according to any one of claims 1-28, wherein A is C(O).

31. The compound according to any one of claims 1-28, wherein A is alkancdiylc s or substituted alkanediylc<8.

32. The compound of claim 31, wherein A is alkanediylc<8-33. The compound of either claim 31 or claim 32, wherein A is methylene or ethylene.

34. The compound according to any one of claims 2-33, wherein m is 1.

35. The compound according to any one of claims 2-33, wherein m is 2.

36. The compound according to any one of claims 2-33, wherein m is 3.

37. The compound according to any one of claims 1-36, wherein n is 1.

38. The compound according to any one of claims 1-36, wherein n is 2.

39. The compound according to any one of claims 1-38, wherein R4 and RT are different.

40. The compound according to any one of claims 1-39, wherein the carbon atom to which R4 and R4' are attached is in the R configuration.

41. The compound according to any one of claims 1-39, wherein the carbon atom to which R4 and R4' are attached is in the S configuration.

42. The compound according to any one of claims 1-41, wherein R4' is hydrogen.

43. The compound according to any one of claims 1-41, wherein R4 is taken together with R4' to form a cycloalkanec<8 or substituted cycloalkanec<8 group.

44. The compound of claim 43, wherein R4 is taken together with R4' to form a cycloalkanec<8 group.

45. The compound of claim 44, wherein the cycloalkanec<8 group is a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, or a cyclohexyl group.

46. The compound according to any one of claims 1-42, wherein R4 is alkylc<8 or substituted alkylc<8-47. The compound of claim 46, wherein R4 is alkylc<8-48. The compound of claim 47, wherein R4 is t-butyl.

49. The compound according to any one of claims 1-42, wherein R4 is aralkylc<8 or substituted aralkylc<8-50. The compound of claim 49, wherein R4 is aralkylc<8-51. The compound of claim 50, wherein R4 is benzyl.

52. The compound according to any one of claims 1-42, wherein R4 is arylc<8 or substituted arylc<8-53. The compound of claim 52, wherein R4 is arylc<8-54. The compound of claim 53, wherein R4 is phenyl.

55. The compound according to any one of claims 1-54, wherein Rb is hydrogen.

56. The compound according to any one of claims 1-55, wherein Rcis hydrogen.

57. The compound according to any one of claims 1-56, wherein the carbon atom to which R5 is attached is in the R configuration.

58. The compound according to any one of claims 1-56, wherein the carbon atom to which R5 is attached is in the S configuration.

59. The compound according to any one of claims 1-4 and 6-58, wherein R5 is hydroxy.

60. The compound according to any one of claims 1-59, wherein the carbon atom to which Re is attached is in the R configuration.

61. The compound according to any one of claims 1-59, wherein the carbon atom to which Re is attached is in the S configuration.

62. The compound according to any one of claims 1-61, wherein Re is hydrogen.

63. The compound according to any one of claims 1-61, wherein Re is alkylc<8-64. The compound of claim 63, wherein Re is methyl.

65. The compound according to any one of claims 1-4 and 6-64, wherein R7 is alkylc<8-66. The compound of claim 65, wherein R7 is methyl.

67. The compound according to any one of claims 1-66, wherein the compound is further defined as:or a pharmaceutically acceptable salt thereof.

68. A pharmaceutical composition comprising:(A) a compound according to any one of claims 1-67; and(B) a pharmaceutically acceptable excipient.

69. The pharmaceutical composition of claim 68, wherein the pharmaceutical composition is formulated for oral administration or administration via injection.

70. The pharmaceutical composition of either claim 68 or claim 69, wherein the pharmaceutical composition formulated as a unit dose.

71. A method of treating a disease or disorder in a patient in need thereof comprising administering to the patient in need thereof a therapeutically effective amount of a compound or composition according to any one of claims 1-70.

72. The method of claim 71, wherein the disease or disorder is cancer.

73. The method of claim 72, wherein the cancer is a carcinoma, sarcoma, lymphoma, leukemia, melanoma, mesothelioma, multiple myeloma, or seminoma.

74. The method of claim 72, wherein the cancer is of the bladder, blood, bone, brain, breast, central nervous system, cervix, colon, endometrium, esophagus, gall bladder, gastrointestinal tract, genitalia, genitourinary tract, head, kidney, larynx, liver, lung, muscle tissue, neck, oral or nasal mucosa, ovary, pancreas, prostate, skin, spleen, small intestine, large intestine, stomach, testicle, or thyroid.

75. The method according to any one of claims 71-74, wherein the cancer comprises a mutated p53 gene.

76. The method according to any one of claims 71-74, wherein the cancer comprises a deleted p53 gene.

77. The method according to any one of claims 71-76, wherein the cancer is breast cancer.

78. The method of claim 77, wherein the breast cancer is triple negative breast cancer.

79. The method according to any one of claims 71-76, wherein the cancer is ovarian cancer.

80. The method according to any one of claims 71-76, wherein the cancer is Burkitt lymphoma.

81. The method according to any one of claims 71-76, wherein the cancer is diffuse large B cell lymphoma.

82. The method of claim 81, wherein the diffuse large B cell lymphoma is subtype ABC.

83. The method of claim 81, wherein the diffuse large B cell lymphoma is subtype GCB.

84. The method according to any one of claims 71-76, wherein the cancer is a B-cell leukemia.

85. The method of claim 84, wherein the B-cell leukemia is B-cell acute lymphocytic leukemia.

86. The method according to any one of claims 71-76, wherein the cancer is a T-cell leukemia.

87. The method of claim 86, wherein the T-cell leukemia is T-cell acute lymphocytic leukemia.

88. The method according to any one of claims 71-76, wherein the cancer is a lung cancer.

89. The method of claim 88, wherein the lung cancer is lung adenocarcinoma.

90. The method according to any one of claims 71-76, wherein the cancer is head and neck cancer.

91. The method according to any one of claims 71-76, wherein the cancer is a sarcoma.

92. The method according to any one of claims 71-91, wherein the method further comprises administering a second therapy.

93. The method of claim 92, wherein the second therapy is surgery, a second chemotherapeutic, radiotherapy, or immunotherapy.

94. The method according to any one of claims 71-93, wherein the compound is administered once.

95. The method according to any one of claims 71-93, wherein the compound is administered two or more times.

96. A method of inhibiting cell replication by modulating mdm2 comprising contacting the cell with a compound or composition according to any one of claims 1-70.

97. A method of modulating the activity of mdm2 in a cell comprising contacting the cell with a compound or composition according to any one of claims 1-70.

98. The method of either claim 96 or claim 97, wherein the method is performed in vitro.

99. The method of either claim 96 or claim 97, wherein the method is performed in vivo.

100. The method of either claim 96 or claim 97, wherein the method is performed ex vivo.

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